Sea Slug Kleptoplasty and Plastid Maintenance in a Metazoan 1[W]

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1 Update on Sea Slug Kleptoplasty and Plastid Maintenance in a Metazoan Sea Slug Kleptoplasty and Plastid Maintenance in a Metazoan 1[W] Karen N. Pelletreau 2, Debashish Bhattacharya 2,DanaC.Price,JaredM.Worful 3, Ahmed Moustafa 4, and Mary E. Rumpho* Department of Molecular and Biomedical Sciences, University of Maine, Orono, Maine (K.N.P., J.M.W., M.E.R.); and Department of Ecology, Evolution, and Natural Resources, Rutgers University, New Brunswick, New Jersey (D.B., D.C.P., A.M.) Trench (1969) was the first to characterize the kleptoplastic (i.e. stolen plastid ) relationship between the sacoglossan mollusc Elysia chlorotica and its algal prey (Vaucheria litorea). In contrast to E. chlorotica, which retains only the plastids of the alga in densely packed digestive tissue (Fig. 1), aquatic invertebrates (e.g. corals, clams, worms, tunicates) and the recently reported spotted salamander (Ambystoma maculatum; work of R. Kerney, reported by Petherick, 2010) owe their photosynthetic capacity to the retention of intact unicellular algae (for review, see Rumpho et al., 2011). Photosynthetic sacoglossans vary in the ability to retain plastids and to maintain their functions. Whereas some of these animals can only utilize transferred photosynthate for several hours before the plastids are degraded, others sustain plastid function for months (for review, see Rumpho et al., 2006, 2011; Händeler et al., 2009; Yamamoto et al., 2009). E. chlorotica exhibits one of the longest time frames for plastid maintenance in the absence of algal food, up to 10 to 12 months in the laboratory (for review, see Rumpho et al., 2011). The obvious question to be asked about this system is how the plastids remain photosynthetically active in the absence of algal nuclei that are presumably required to furnish transcripts for plastid-targeted proteins involved in photosynthesis, signaling, regulation, and protein turnover. This update will compare and contrast past approaches used to understand the basis of plastid maintenance and function with recent work using next-generation sequencing to reconcile what appear to be contradictory outcomes in the observed data. 1 This work was supported by the National Science Foundation (grant nos. IOS to M.E.R. and to D.B.) and the National Institutes of Health (grant no. R01ES to D.B.). This is Maine Agricultural and Forest Experiment Station Publication no. 3170, Hatch Project no. ME MRF (NC 1168). 2 These authors contributed equally to the article. 3 Present address: ImmunoGen, Inc., Waltham, MA Present address: Department of Biology and Graduate Program in Biotechnology, American University in Cairo, New Cairo 11835, Egypt. * Corresponding author; mrumpho@umit.maine.edu. [W] The online version of this article contains Web-only data. FUNCTIONAL SEA SLUG KLEPTOPLASTY The exploitation of photosynthesis by heterotrophic organisms is well documented in aquatic (Trench, 1993; Venn et al., 2008) and terrestrial (Nash, 2008) ecosystems. In these opportunistic relationships, the algal symbiont gains refuge and a stable nutrient source in exchange for supplying the host (e.g. invertebrate, amphibian, plant) with carbon (Trench, 1993; Yellowlees et al., 2008). Symbiotic sacoglossans develop a similar relationship, although exploiting only the plastid captured from specific algal prey (Jensen, 1982; Marin and Ros, 2004; Händeler et al., 2009). This phenomenon was recognized in Elysia species by Kawaguti and Yamasu (1965) and Trench (1969), followed by a description of the ecology (Hinde and Smith, 1974; Jensen, 1986; Clark et al., 1990) and development (Harrigan and Alkon, 1978; West, 1979; West et al., 1984). E. chlorotica has an obligate relationship with Vaucheria species, feeding only on Vaucheria litorea (Fig. 1) or Vaucheria compacta (West et al., 1984). Development is predominantly planktonic, and the deposited eggs and planktonic larval veligers lack plastids. Growth of the veligers occurs by feeding on microalgae in the environment, and metamorphosis occurs after 10 to 21 d in the water column. However, settlement and metamorphosis of the veligers into adult sea slugs require the presence of Vaucheria. Most often, the veligers settle on the algal filaments, ensuring a food supply for recently metamorphosed juveniles (West, 1979). Growth and maturation of E. chlorotica are dependent on feeding on Vaucheria for about 1 week, after which plastids are able to support continued growth of the animal (Rumpho et al., 2011). The mechanisms that allow long-term plastid photosynthetic ability in a heterotrophic host have been studied in detail but remain enigmatic. HYPOTHESES TO EXPLAIN THE ENIGMA Retention of Algal Nuclei A reasonable explanation for long-term plastid maintenance is the presence of algal nuclei within the animal that provide the transcripts needed to support plastid functions. Analyses of adult, green, kleptoplastic sacoglossans using microscopy, PCR, Plant Physiology Ò, April 2011, Vol. 155, pp , Ó 2011 American Society of Plant Biologists 1561 Downloaded from on December 12, Published by Copyright 2011 American Society of Plant Biologists. All rights reserved.

2 Pelletreau et al. Figure 1. A, Adult E. chlorotica feeding on its algal prey, the coenocytic heterokont V. litorea. Bar = 5 mm. B, Confocal micrograph showing the red autofluorescent plastids densely packed within the digestive diverticula of E. chlorotica. Once established, these plastids remain photosynthetically active within the adult for several months. Bar = 50 mm. and Southern-blot analysis, however, have failed to substantiate this hypothesis (Graves et al., 1979; Mujer et al., 1996; Green et al., 2000; Rumpho et al., 2001; Mondy and Pierce, 2003; Pierce et al., 2003; Wägele et al., 2011). In addition, molecular markers for nucleusencoded algal genes (e.g. internal transcribed spacer 1 and spermidine synthase) are not found with PCR using DNA derived from starved animal tissue (Pierce et al., 2007, 2009; Rumpho et al., 2008; Schwartz et al., 2010). Plastid Genetic Autonomy Another potential explanation for long-term plastid function in E. chlorotica is the existence of a genetically autonomous V. litorea plastid genome. It is formally possible that this genome has regained, via horizontal gene transfer (HGT), critical genes encoding plastid proteins involved in photosynthesis that have been transferred to the nucleus in other algae and plants. To address this idea, Rumpho et al. (2008) sequenced the plastid genome of V. litorea and found it to be comparable to other algal and plant plastids in terms of gene content and found no evidence for HGT. Plastid Stability Although containing a typical algal genome, the Vaucheria plastid exhibits unique physical and biochemical properties that may play a role in the establishment of the sacoglossan symbiosis (Jensen, 1982; Händeler et al., 2009; Wägele et al., 2011). Vaucheria, like many of the algae consumed by sacoglossan molluscs, is filamentous and coenocytic: in essence, a single multinucleate cell. This allows the sea slug to rapidly acquire numerous plastids while feeding but also favors plastids that appear to have a greater longevity than those associated with multicellular algae. Green et al. (2005) investigated the physiology of isolated Vaucheria plastids and found that 30% to 40% (in light versus dark, respectively) remained intact 14 d after isolation from the alga, whereas less than 20% of isolated spinach (Spinacia oleracea) chloroplasts were intact after 24 h. Isolated V. litorea plastids exhibited electron transport activity, CO 2 -dependent oxygen evolution, and CO 2 fixation 72 h post isolation. The isolated plastids were also largely unaffected by osmotic fluctuations, tested up to 670 mm mannitol. In addition, over a 3-d period, de novo synthesis of plastid proteins in isolated plastids changed minimally in banding patterns and intensity on SDS-PAGE gels, and both Rubisco (large and small subunits) and the PSII D1 protein were synthesized in plastids 3 d post isolation. These data support the unique nature of V. litorea plastids in not requiring s-factors or other nucleus-encoded regulatory factors for transcription and translation of plastid genes or that the factors are stable for a minimum of 3 d in vitro. Dual Targeting of Proteins Dual targeting of animal-derived proteins as an explanation for plastid replenishment has yet to be carefully studied. All of the nucleus-encoded enzymes of the Calvin-Benson carbon reduction cycle have cytosolic counterparts in E. chlorotica except for phosphoribulokinase and sedoheptulose-1,7-bisphosphatase (both subunits of Rubisco are plastid encoded in V. litorea). In addition, transcripts for all of these nucleusencoded enzymes are present in the partial E. chlorotica transcriptome library discussed in more detail below (Supplemental Table S1). Because these proteins are encoded in the animal genome, they could be coopted for use in the plastid, providing much of the necessary machinery for carbon fixation. HGT The most highly cited hypothesis to date, and the focus of this update, is extensive HGT from the algal nucleus to the animal, thereby supporting long-term plastid function (Olendzenski and Gogarten, 2009; Bock, 2010; Boto, 2010; Moran and Jarvik, 2010). The majority of the existing work (Table I; see refs. below) has investigated the presence of genes required for photosynthesis in the nuclear genome and transcriptome of E. chlorotica in both adults and larvae (with and without plastids present, respectively). Until recently, these studies used the candidate gene approach to putatively identify alga-derived genes in the animal. The methods have included PCR (Green et al., 2000; Mondy and Pierce, 2003; Pierce et al., 2003, 2007, 1562 Plant Physiol. Vol. 155, 2011 Downloaded from on December 12, Published by Copyright 2011 American Society of Plant Biologists. All rights reserved.

3 Plastid Maintenance in a Metazoan Table I. Molecular and biochemical evidence for nucleus-encoded algal genes in E. chlorotica Protein Method Gene Source Fucoxanthin a/c chlorophyll-binding protein 35 S immunolabeling fcp Pierce et al. (1996, 2003) Fucoxanthin a/c chlorophyll-binding protein PCR fcp Pierce et al. (2007) Light-harvesting complex-binding protein 1 35 S immunolabeling lhcv-1 Hanten and Pierce (2001) Light-harvesting complex-binding protein 1 PCR lhcv-1 Pierce et al. (2007) Light-harvesting complex-binding protein 2 PCR lhcv-2 Pierce et al. (2007) Light-harvesting complex-binding protein 3 PCR lhcv-3 Pierce et al. (2009) Light-harvesting complex-binding protein 4 PCR lhcv-4 Pierce et al. (2009) Oxygen evolution complex protein PCR psbo Rumpho et al. (2008) Oxygen evolution complex protein Quantitative PCR psbo K. Soule and M.E. Rumpho (unpublished data) Phosphoribulokinase 35 S immunolabeling prk Rumpho et al. (2009) Phosphoribulokinase PCR prk Rumpho et al. (2009) Phosphoribulokinase Quantitative PCR prk Soule (2009) Uroporphyrinogen decarboxylase PCR urod Pierce et al. (2009) a Magnesium chelatase subunit D PCR chld Pierce et al. (2009) a Magnesium chelatase subunit H PCR chlh Pierce et al. (2009) a Chlorophyll synthase PCR chlg Pierce et al. (2009) a Chlorophyll synthesis 14 C labeling ND b Pierce et al. (2009) a Where identical data were presented in more than one publication, the earliest publication date is noted. b ND, Not determined. 2009; Rumpho et al., 2008, 2009), detection of de novo synthesis of nucleus-encoded plastid proteins using radiolabeling and immunolabeling in combination with specific inhibitors of transcription or translation (Pierce et al., 1996, 2007; Green et al., 2000; Hanten and Pierce, 2001; Rumpho et al., 2001, 2009), northern-blot analysis and genome walking (Rumpho et al., 2008), 14 C incorporation and synthesis of chlorophyll in animal tissue (Pierce et al., 2009), and quantitative reverse transcription-pcr of nucleus-encoded algal genes in the animal (Soule, 2009). All of these data support the presence of a few (Rumpho et al., 2008, 2009) to numerous (Pierce et al., 2009; Schwartz et al., 2010) nuclear algal genes or gene fragments in E. chlorotica. Recently, partial transcriptome data from two kleptoplastic sea slugs, Elysia timida and Plakobranchus ocellatus, were generated by Wägele et al. (2011). Both of these sea slugs feed on multiple chlorophycean algae in nature and retain functional plastids for about 57 and 69 d, respectively (Evertsen et al., 2007). Analysis of 77,648 ESTs from a single P. ocellatus individual and 24,200 ESTs from 15 E. timida individuals showed that 96% to 98% of the expressed genes were metazoan and none were of algal provenance. Based on these data, the authors concluded that HGT plays no role in sustaining long-term plastid function in these sea slugs. Rather, they attributed this unique relationship to the physical characteristics of the algal plastids and the morphology and cellular environment of the animal (physically and biochemically protecting the plastids), based on their ultrastructural, pulse amplitude modulated fluorescence, and phylogenetic analyses (Wägele and Johnsen, 2001; Evertsen et al., 2007; Evertsen and Johnsen, 2009; Händeler et al., 2009). Using essentially the same approach, we generated partial transcriptome data from E. chlorotica to investigate the extent of HGT within this system (Supplemental Materials and Methods S1; Supplemental Table S1; to download a complete list of the isotigs and singletons produced by GS Assembler, see dblab.rutgers.edu/home/downloads/). Unlike the sacoglossans studied by Wägele et al. (2011), cumulative data (summarized above and in Table I) resulting from multiple experimental approaches and investigators over many years have supported HGT in the E. chlorotica/v. litorea system, although the extent of this transfer has been in question: in several key genes (Rumpho et al., 2008, 2009) or in entire metabolic pathways (Pierce et al., 2009; Schwartz et al., 2010). Using 454 pyrosequencing, we generated 148 Mb of cdna sequence data from starved but actively photosynthesizing adult E. chlorotica (n = 5). From this partial transcriptome, 13,978 assembled unigenes and 99,873 unassembled singletons were analyzed using BLASTx (e-value cutoff # ) to generate putative gene annotations and to assign the taxonomic origin of ESTs. As expected, at least 95% of the predicted proteins had top hits to Metazoa. The putative nonmetazoan top hits returned by BLASTx analysis of the E. chlorotica unigenes (123) and singletons (354) were used as queries in a phylogenomic pipeline specifically designed to identify genes of foreign origin from the host using the methods and database described by Moustafa et al. (2009). This approach identified 20 ESTs of potential foreign origin derived from different prokaryotes, eukaryotes, and viruses (Supplemental Table S2) as well as several plastid-derived transcripts primarily from V. litorea, indicating plastid activity (Supplemental Table S3). None of these 20 ESTs, however, has a direct involvement in photosynthesis. In addition, specific BLASTn searches of both the contig and singleton data for genes previously identified as HGT candidates (lhcv-1, -2, -3, and -4; fcp; psbo; prk; Plant Physiol. Vol. 155, Downloaded from on December 12, Published by Copyright 2011 American Society of Plant Biologists. All rights reserved.

4 Pelletreau et al. urod; chld, -H, and -G; Table I) failed to identify homologs. In all of these transcriptome studies, no evidence exists to indicate that nucleus-encoded algal genes are expressed in photosynthetic sacoglossans. Wägele et al. (2011) noted the high expression level of photosynthetic genes in the transcriptome of the source alga Acetabularia, comparable to that observed in the model green plant Arabidopsis (Arabidopsis thaliana), and yet none of the top 50 highly expressed nuclear transcripts for plastid protein were found in the two sacoglossans used in their study. Although data are not publicly available from the V. litorea transcriptome to conduct such a comparative analysis (Pierce et al., 2009), one can assume that a similar hierarchy of expression patterns would be observed in this alga. In the partial transcriptome data presented here from E. chlorotica, we also did not find evidence of expression of any of the photosynthetic top 50 nucleus-encoded algal genes. However, it is important to note that expression patterns of a putatively transferred gene in the animal may not mirror what is observed in the alga, and to assume similar expression patterns in the foreign environment is highly speculative. Not only might the copy number of the genes be markedly different, but the expression levels and modes of regulation of expression in the animal are unknown. Recent studies employing quantitative reverse transcription-pcr support the expression of both prk and psbo in starved E. chlorotica. However, the expression levels of these genes in the animal were markedly lower, as much as 525 times lower for prk (Soule, 2009) and 63 times lower for psbo (K. Soule and M.E. Rumpho, unpublished data), compared with expression levels measured in V. litorea under identical (2 h post illumination) conditions. Differential expression of both prk and psbo in the animal was observed over a diurnal cycle, but again these patterns were markedly different from that observed in V. litorea. Similar patterns were also observed with the plastid-encoded transcripts rbcl and psaa. These studies provide evidence of the expression of nucleus-encoded plastidtargeted proteins in E. chlorotica starved for several months, but more importantly, they suggest that levels and patterns of expression of foreign genes may not mirror those in the alga (e.g. Acetabularia or Vaucheria). Thus, although Wägele et al. (2011) provide compelling arguments to suggest that nucleus-encoded algal transcripts could not have been missed in their transcriptome libraries, we feel that more exhaustive sequencing may be required to adequately address this issue. Of interest in both studies is the finding of plastid gene expression, indicating that this genome is actively transcribed in the animals. The 46 plastid ESTs identified included 19 unique genes in E. chlorotica, some of which were represented multiple times (Supplemental Table S3). Together with the data from Wägele et al. (2011), these data suggest that nucleusencoded transcription factors or regulatory molecules are not necessarily required for plastid gene expression in these taxa. FUTURE DIRECTIONS Alternative hypotheses to HGT need to be considered to explain plastid function in sacoglossans. It is apparent that plastid stability is essential for the success of these symbioses, from initial uptake to establishment of kleptoplasty (Green et al., 2005; Wägele et al., 2011). The extent to which HGT is occurring is less clear and warrants further exploration to resolve the conflicting results to date. Wägele et al. (2011) conclude that HGT does not explain plastid longevity and function in the two sacoglossans investigated in their study. A similar conclusion cannot yet be drawn for E. chlorotica due to the longevity of this functional association and multiple lines of evidence that indicate that nuclear algal genes have been transferred. Therefore, either these sacoglossan species (E. chlorotica versus E. timida and P. ocellatus) have evolved unique mechanisms to obtain and sustain photosynthetic abilities, one utilizing gene transfer and the others not, or additional explanations are required to reconcile current data. Finally, reconciliation of these data will only occur through the investigation of new mechanisms that could be working synergistically with limited HGT and plastid stability toward plastid function. This could include investigating whether (1) the animal cells transiently use proteins and transcripts in an opportunistic fashion when available, (2) transcripts are integrated into extrachromosomal elements, which may restrict detection due to extremely low copy number and expression levels, (3) the unique stability of the plastids translates to unusual protein turnover rates of essential plastid- and nucleus-encoded photosynthetic proteins in the animal versus alga and/or protease activity, and (4) the regulation of expression of foreign genes in the animal impacts traditional detection approaches. Supplemental Data The following materials are available in the online version of this article. Supplemental Table S1. List of Metazoa-derived unigenes. Supplemental Table S2. ESTs of putative nonmetazoan origin. Supplemental Table S3. ESTs of plastid origin. Supplemental Materials and Methods S1. Library preparation and bioinformatic analysis. Received February 8, 2011; accepted February 20, 2011; published February 23, LITERATURE CITED Bock R (2010) The give-and-take of DNA: horizontal gene transfer in plants. Trends Plant Sci 15: Boto L (2010) Horizontal gene transfer in evolution: facts and challenges. Proc Biol Sci 277: Plant Physiol. Vol. 155, 2011 Downloaded from on December 12, Published by Copyright 2011 American Society of Plant Biologists. All rights reserved.

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6 Supplemental Methods Preparation and sequencing of cdna library Elysia chlorotica Gould was collected from Martha s Vineyard, MA, in October Animals were maintained in laboratory culture at 10 C under a 12:12 L:D cycle in artificial seawater (ASW; Instant Ocean) at 32 PSU. The animals were starved (i.e., deprived of algal food) for 5 weeks before RNA extraction. Five individual sea slugs were placed in the dark for 72 h, illuminated for 3 h and then flash frozen in liquid nitrogen. RNA was extracted using Trizol (Invitrogen) followed by polya selected SMART cdna synthesis and normalization (Moscow, Russia; (Zhu et al., 2001; Zhulidov et al., 2004). Normalized, double stranded cdna was validated using PCR amplification and threshold crossing between a common (E. chlorotica actin) and rare (Vaucheria litorea plastid encoded psbb) gene product. Normalized cdna was sequenced using 454-pyrosequencing technology at the Research Technology Support Facility at Michigan State University in October 2007, and subsequently at the Roy J. Carver Center for Comparative Genomics at the University of Iowa in March The two pyrosequencing runs resulted in 619,612 reads of high quality totaling 147,802,132 bp of data. These sequences were co-assembled as cdna using the Roche GS De Novo Assembler V2.3. The assembly contained 13,978 unigenes (contigs/isotigs) with an average size of 739 bp with a N50 size of 931 bp. The largest contig was 4,958 bp in size. A total of 99,873 reads could not be assembled (i.e., were singletons). The isotigs and singletons produced by the GS Assembler are available for download at Protein Prediction The top hit for each EST unigene and singleton was determined using BLASTx searches (e-value cut-off <10-10 ) against a database of annotated proteins extracted from GenBank ( the Joint Genome Institute (JGI; as well as six-frame protein translations of EST sequences from algal and microbial eukaryotes extracted from dbest at GenBank, TBestDB, EST data from Porphyridium cruentum and partial genome data from Calliarthron tuberculosum (the latter two to enrich the database of red algal genomic data). Metazoan and non-metazoan top hits were separated allowing us to focus the phylogenomic analysis on a reduced set of putative foreign genes to test for the presence of transferred genes in the animal. We generated putative annotations for both metazoan and non-metazoan sequences by transferring the BLASTx top hit

7 annotation to each of them. A key set of ESTs that were inspected here were plastid-encoded genes most likely generated as artefacts because our cdna library preparation procedure was designed to exclude organelle mrnas with the use of polya selection. Regardless, these ESTs provide evidence of plastid gene expression in the starved sea slugs. Phylogenomic analysis The non-metazoan top hits that were returned by BLASTx analysis of the E. chlorotica unigenes and singletons were used as queries in our local phylogenomic pipeline with the local database described above (e.g. Moustafa et al., 2009). Here, each group of homologous protein sequences identified by BLAST were aligned using MAFFT (Katoh et al., 2005), and molecular phylogenies were inferred using maximum likelihood (RAxML v7.0.4, Stamatakis, 2006) under the WAG amino acid substitution model (Whelan and Goldman, 2001) and a discrete gamma distribution (Yang, 1994). Branch support was evaluated with 100 bootstrap replicates using RAxML. Thereafter, we manually inspected each tree to determine whether they still provided support for the HGT model or were false positives resulting from the BLASTx top-hit approach. Katoh K, Kuma K, Toh H, Miyata T (2005) MAFFT version 5: improvement in accuracy of multiple sequence alignment. Nucleic Acids Res 33: Moustafa A, Beszteri B, Maier UG, Bowler C, Valentin K, Bhattacharya D (2009) Genomic footprints of a cryptic plastid endosymbiosis in diatoms. Science 324: Stamatakis A (2006) RAxML-VI-HPC: Maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed models. Bioinformatics 22: Whelan S, Goldman N (2001) A general empirical model of protein evolution derived from multiple protein families using a maximum-likelihood approach. Mol Biol Evol 18: Yang ZH (1994) Maximum likelihood phylogenetic estimation from DNA sequences with variable rates over sites - approximate methods. J Mol Evol 39:

8 Zhu YY, Machleder EM, Chenchik A, Li R, Siebert PD (2001) Reverse transcriptase template switching: A SMART (TM) approach for full-length cdna library construction. Biotechniques 30: Zhulidov PA, Bogdanova EA, Shcheglov AS, Vagner LL, Khaspekov GL, Kozhemyako VB, Matz MV, Meleshkevitch E, Moroz LL, Lukyanov SA, Shagin DA (2004) Simple cdna normalization using kamchatka crab duplex-specific nuclease. Nucleic Acids Res 32: 15

9 Table S1. List of metazoan-derived unigenes and their top hits found using BLASTx and our comprehensive local database. The gene ontology (GO) annotations are also presented. SeqName Hit-Desc GO-Group GO-ID Term Organism e-val isotig00031 mmset type ii molecular_functiongo: binding Branchiostoma floridae 2.10E-29 isotig00031 mmset type ii molecular_functiongo: transferase activity Branchiostoma floridae 2.10E-29 contig00226leucyl-trna synthetase molecular_functiongo: ligase activity Ciona intestinalis 9.48E-86 contig00226leucyl-trna synthetase biological_process GO: translation Ciona intestinalis 9.48E-86 isotig00117 tenascin r ( janusin) biological_process GO: regulation of cellular process Pan troglodytes 3.05E-48 isotig00117 tenascin r ( janusin) biological_process GO: cellular component organization Pan troglodytes 3.05E-48 isotig00118 tenascin r ( janusin) biological_process GO: axon guidance Pan troglodytes 9.52E-46 isotig00118 tenascin r ( janusin) biological_process GO: regulation of cellular process Pan troglodytes 9.52E-46 isotig00118 tenascin r ( janusin) cellular_componen GO: cell part Pan troglodytes 9.52E-46 isotig00118 tenascin r ( janusin) cellular_componen GO: extracellular region Pan troglodytes 9.52E-46 isotig00119 tenascin r ( janusin) biological_process GO: cellular process Callithrix jacchus 4.41E-42 isotig00119 tenascin r ( janusin) biological_process GO: cellular component organization Callithrix jacchus 4.41E-42 isotig00120 tenascin r ( janusin) biological_process GO: cellular process Biomphalaria glabrata 8.96E-52 isotig00120 tenascin r ( janusin) cellular_componen GO: extracellular region Biomphalaria glabrata 8.96E-52 isotig00120 tenascin r ( janusin) biological_process GO: cellular component organization Biomphalaria glabrata 8.96E-52 isotig00121 tenascin r ( janusin) biological_process GO: cellular process Pongo abelii 1.06E-48 isotig00121 tenascin r ( janusin) biological_process GO: cellular component organization Pongo abelii 1.06E-48 isotig00122 tenascin r ( janusin) biological_process GO: cellular process Pan troglodytes 3.97E-51 isotig00123 tenascin r ( janusin) biological_process GO: cellular process Biomphalaria glabrata 1.28E-51 isotig00123 tenascin r ( janusin) cellular_componen GO: extracellular region Biomphalaria glabrata 1.28E-51 isotig00124 tenascin r ( janusin) biological_process GO: cellular process Callithrix jacchus 1.93E-52 isotig00124 tenascin r ( janusin) biological_process GO: cellular component organization Callithrix jacchus 1.93E-52 isotig00125 tenascin r ( janusin) biological_process GO: cellular process Callithrix jacchus 1.95E-52 isotig00138 l _3-like molecular_functiongo: nucleic acid binding Danio rerio 3.51E-57 isotig00138 l _3-like biological_process GO: nucleobase, nucleoside, nucleotide and nucleic acid metabolic process Danio rerio 3.51E-57 isotig00139 l _3-like molecular_functiongo: nucleic acid binding Danio rerio 3.84E-57 isotig00139 l _3-like biological_process GO: nucleobase, nucleoside, nucleotide and nucleic acid metabolic process Danio rerio 3.84E-57 isotig00140 l _3-like molecular_functiongo: nucleic acid binding Danio rerio 3.55E-57 isotig00140 l _3-like biological_process GO: nucleobase, nucleoside, nucleotide and nucleic acid metabolic process Danio rerio 3.55E-57 isotig00141 l _3-like molecular_functiongo: nucleic acid binding Danio rerio 3.89E-57 isotig00141 l _3-like biological_process GO: nucleobase, nucleoside, nucleotide and nucleic acid metabolic process Danio rerio 3.89E-57 isotig00146 l _3-like molecular_functiongo: nucleic acid binding Danio rerio 4.37E-57 isotig00146 l _3-like biological_process GO: nucleobase, nucleoside, nucleotide and nucleic acid metabolic process Danio rerio 4.37E-57 isotig00148 l _3-like molecular_functiongo: nucleic acid binding Danio rerio 3.89E-57 isotig00148 l _3-like biological_process GO: nucleobase, nucleoside, nucleotide and nucleic acid metabolic process Danio rerio 3.89E-57 isotig00149 l _3-like molecular_functiongo: nucleic acid binding Danio rerio 3.85E-57 isotig00149 l _3-like biological_process GO: nucleobase, nucleoside, nucleotide and nucleic acid metabolic process Danio rerio 3.85E-57 isotig00150 l _3-like molecular_functiongo: nucleic acid binding Danio rerio 3.51E-57 isotig00150 l _3-like biological_process GO: nucleobase, nucleoside, nucleotide and nucleic acid metabolic process Danio rerio 3.51E-57 isotig00151 l _3-like molecular_functiongo: nucleic acid binding Danio rerio 4.07E-57 isotig00151 l _3-like biological_process GO: nucleobase, nucleoside, nucleotide and nucleic acid metabolic process Danio rerio 4.07E-57 isotig00152 l _3-like molecular_functiongo: nucleic acid binding Danio rerio 3.55E-57 isotig00152 l _3-like biological_process GO: nucleobase, nucleoside, nucleotide and nucleic acid metabolic process Danio rerio 3.55E-57 isotig00153 l _3-like molecular_functiongo: nucleic acid binding Danio rerio 4.37E-57 isotig00153 l _3-like biological_process GO: nucleobase, nucleoside, nucleotide and nucleic acid metabolic process Danio rerio 4.37E-57 isotig00154 l _3-like molecular_functiongo: nucleic acid binding Danio rerio 4.00E-57 isotig00154 l _3-like biological_process GO: nucleobase, nucleoside, nucleotide and nucleic acid metabolic process Danio rerio 4.00E-57 isotig00156 l _3-like molecular_functiongo: nucleic acid binding Danio rerio 4.08E-57 isotig00156 l _3-like biological_process GO: nucleobase, nucleoside, nucleotide and nucleic acid metabolic process Danio rerio 4.08E-57 isotig00162 mitogen-activated protein kinase 8 molecular_functiongo: jun kinase activity Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 cellular_componen GO: nucleoplasm Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: response to hydrogen peroxide Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 molecular_functiongo: calmodulin-dependent protein kinase activity Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: peptidyl-threonine phosphorylation Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 cellular_componen GO: vesicle Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: regulation of transcription, DNA-dependent Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: JUN phosphorylation Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: ossification Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: response to steroid hormone stimulus Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: neuron projection development Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: response to UV Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 cellular_componen GO: cytosol Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: response to osmotic stress Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 cellular_componen GO: dendrite cytoplasm Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: induction of apoptosis in response to chemical stimulus Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: inflammatory response Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: response to cadmium ion Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: response to heat Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: activation of pro-apoptotic gene products Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 cellular_componen GO: axon part Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: positive regulation of DNA replication Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: positive regulation of microtubule polymerization Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 molecular_functiongo: calmodulin binding Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: positive regulation of cell migration Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 molecular_functiongo: atp binding Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 biological_process GO: negative regulation of apoptosis Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 cellular_componen GO: perikaryon Macaca mulatta 1.37E-17 isotig00162 mitogen-activated protein kinase 8 cellular_componen GO: mitochondrion Macaca mulatta 1.37E-17 isotig00163 mitogen-activated protein kinase 8 molecular_functiongo: jun kinase activity Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 cellular_componen GO: nucleoplasm Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: response to hydrogen peroxide Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 molecular_functiongo: calmodulin-dependent protein kinase activity Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: peptidyl-threonine phosphorylation Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 cellular_componen GO: vesicle Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: regulation of transcription, DNA-dependent Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: JUN phosphorylation Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: ossification Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: response to steroid hormone stimulus Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: neuron projection development Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: response to UV Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 cellular_componen GO: cytosol Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: response to osmotic stress Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 cellular_componen GO: dendrite cytoplasm Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: induction of apoptosis in response to chemical stimulus Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: inflammatory response Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: response to cadmium ion Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: response to heat Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: activation of pro-apoptotic gene products Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 cellular_componen GO: axon part Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: positive regulation of DNA replication Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: positive regulation of microtubule polymerization Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 molecular_functiongo: calmodulin binding Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: positive regulation of cell migration Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 molecular_functiongo: atp binding Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 biological_process GO: negative regulation of apoptosis Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 cellular_componen GO: perikaryon Macaca mulatta 1.30E-17 isotig00163 mitogen-activated protein kinase 8 cellular_componen GO: mitochondrion Macaca mulatta 1.30E-17 isotig00171 palmitoylated membrane protein 2 isoform 2 molecular_functiongo: guanylate kinase activity Branchiostoma floridae 1.64E-24 isotig00171 palmitoylated membrane protein 2 isoform 2 cellular_componen GO: membrane fraction Branchiostoma floridae 1.64E-24 isotig00171 palmitoylated membrane protein 2 isoform 2 cellular_componen GO: cell surface Branchiostoma floridae 1.64E-24 isotig00171 palmitoylated membrane protein 2 isoform 2 biological_process GO: signal transduction Branchiostoma floridae 1.64E-24 isotig00171 palmitoylated membrane protein 2 isoform 2 cellular_componen GO: integral to plasma membrane Branchiostoma floridae 1.64E-24 isotig00171 palmitoylated membrane protein 2 isoform 2 molecular_functiongo: protein binding Branchiostoma floridae 1.64E-24 isotig00173 palmitoylated membrane protein 2 isoform 2 molecular_functiongo: guanylate kinase activity Branchiostoma floridae 1.85E-24 isotig00173 palmitoylated membrane protein 2 isoform 2 cellular_componen GO: membrane fraction Branchiostoma floridae 1.85E-24 isotig00173 palmitoylated membrane protein 2 isoform 2 cellular_componen GO: cell surface Branchiostoma floridae 1.85E-24 isotig00173 palmitoylated membrane protein 2 isoform 2 biological_process GO: signal transduction Branchiostoma floridae 1.85E-24 isotig00173 palmitoylated membrane protein 2 isoform 2 cellular_componen GO: integral to plasma membrane Branchiostoma floridae 1.85E-24 isotig00173 palmitoylated membrane protein 2 isoform 2 molecular_functiongo: protein binding Branchiostoma floridae 1.85E-24 isotig00175 palmitoylated membrane protein 2 isoform 2 molecular_functiongo: guanylate kinase activity Branchiostoma floridae 1.64E-24 isotig00175 palmitoylated membrane protein 2 isoform 2 cellular_componen GO: membrane fraction Branchiostoma floridae 1.64E-24

10 isotig00175 palmitoylated membrane protein 2 isoform 2 cellular_componen GO: cell surface Branchiostoma floridae 1.64E-24 isotig00175 palmitoylated membrane protein 2 isoform 2 biological_process GO: signal transduction Branchiostoma floridae 1.64E-24 isotig00175 palmitoylated membrane protein 2 isoform 2 cellular_componen GO: integral to plasma membrane Branchiostoma floridae 1.64E-24 isotig00175 palmitoylated membrane protein 2 isoform 2 molecular_functiongo: protein binding Branchiostoma floridae 1.64E-24 isotig00183 palmitoylated membrane protein 2 isoform 2 molecular_functiongo: guanylate kinase activity Branchiostoma floridae 4.60E-13 isotig00183 palmitoylated membrane protein 2 isoform 2 cellular_componen GO: membrane fraction Branchiostoma floridae 4.60E-13 isotig00183 palmitoylated membrane protein 2 isoform 2 cellular_componen GO: cell surface Branchiostoma floridae 4.60E-13 isotig00183 palmitoylated membrane protein 2 isoform 2 biological_process GO: signal transduction Branchiostoma floridae 4.60E-13 isotig00183 palmitoylated membrane protein 2 isoform 2 cellular_componen GO: integral to plasma membrane Branchiostoma floridae 4.60E-13 isotig00183 palmitoylated membrane protein 2 isoform 2 molecular_functiongo: protein binding Branchiostoma floridae 4.60E-13 isotig00249 endonuclease-reverse transcriptase molecular_functiongo: catalytic activity Strongylocentrotus purpuratus 6.14E-10 isotig00260 tenascin r ( janusin) biological_process GO: cellular process Biomphalaria glabrata 2.44E-51 isotig00261 tenascin r cellular_componen GO: cell surface Biomphalaria glabrata 2.30E-49 isotig00261 tenascin r molecular_functiongo: binding Biomphalaria glabrata 2.30E-49 isotig00261 tenascin r biological_process GO: extracellular matrix organization Biomphalaria glabrata 2.30E-49 isotig00261 tenascin r cellular_componen GO: membrane raft Biomphalaria glabrata 2.30E-49 isotig00262 tenascin r ( janusin) biological_process GO: cellular process Biomphalaria glabrata 7.56E-53 isotig00265 tenascin r ( janusin) biological_process GO: axon guidance Biomphalaria glabrata 7.21E-54 isotig00265 tenascin r ( janusin) cellular_componen GO: extracellular region Biomphalaria glabrata 7.21E-54 isotig00266 tenascin r ( janusin) biological_process GO: axon guidance Biomphalaria glabrata 9.27E-54 isotig00266 tenascin r ( janusin) cellular_componen GO: extracellular region Biomphalaria glabrata 9.27E-54 isotig00267 tenascin r ( janusin) biological_process GO: cellular process Biomphalaria glabrata 1.05E-54 isotig00267 tenascin r ( janusin) cellular_componen GO: extracellular region Biomphalaria glabrata 1.05E-54 isotig00267 tenascin r ( janusin) biological_process GO: cellular component organization Biomphalaria glabrata 1.05E-54 isotig ecto-like molecular_functiongo: binding Saccoglossus kowalevskii 1.02E-63 isotig ecto-like biological_process GO: nucleobase, nucleoside and nucleotide metabolic process Saccoglossus kowalevskii 1.02E-63 isotig ecto-like molecular_functiongo: '-nucleotidase activity Saccoglossus kowalevskii 1.02E-63 isotig ecto-like cellular_componen GO: cytoplasm Saccoglossus kowalevskii 1.02E-63 isotig ecto-like biological_process GO: cellular metabolic process Saccoglossus kowalevskii 1.02E-63 isotig ecto-like cellular_componen GO: plasma membrane Saccoglossus kowalevskii 1.02E-63 isotig ecto-like molecular_functiongo: binding Saccoglossus kowalevskii 3.31E-59 isotig ecto-like biological_process GO: nucleobase, nucleoside and nucleotide metabolic process Saccoglossus kowalevskii 3.31E-59 isotig ecto-like molecular_functiongo: '-nucleotidase activity Saccoglossus kowalevskii 3.31E-59 isotig ecto-like cellular_componen GO: cytoplasm Saccoglossus kowalevskii 3.31E-59 isotig ecto-like biological_process GO: cellular metabolic process Saccoglossus kowalevskii 3.31E-59 isotig ecto-like cellular_componen GO: plasma membrane Saccoglossus kowalevskii 3.31E-59 isotig ecto-like molecular_functiongo: binding Saccoglossus kowalevskii 1.03E-63 isotig ecto-like biological_process GO: nucleobase, nucleoside and nucleotide metabolic process Saccoglossus kowalevskii 1.03E-63 isotig ecto-like molecular_functiongo: '-nucleotidase activity Saccoglossus kowalevskii 1.03E-63 isotig ecto-like cellular_componen GO: cytoplasm Saccoglossus kowalevskii 1.03E-63 isotig ecto-like biological_process GO: cellular metabolic process Saccoglossus kowalevskii 1.03E-63 isotig ecto-like cellular_componen GO: plasma membrane Saccoglossus kowalevskii 1.03E-63 isotig ecto-like molecular_functiongo: binding Saccoglossus kowalevskii 3.32E-59 isotig ecto-like biological_process GO: nucleobase, nucleoside and nucleotide metabolic process Saccoglossus kowalevskii 3.32E-59 isotig ecto-like molecular_functiongo: '-nucleotidase activity Saccoglossus kowalevskii 3.32E-59 isotig ecto-like cellular_componen GO: cytoplasm Saccoglossus kowalevskii 3.32E-59 isotig ecto-like biological_process GO: cellular metabolic process Saccoglossus kowalevskii 3.32E-59 isotig ecto-like cellular_componen GO: plasma membrane Saccoglossus kowalevskii 3.32E-59 isotig ecto-like molecular_functiongo: binding Saccoglossus kowalevskii 9.76E-64 isotig ecto-like biological_process GO: nucleobase, nucleoside and nucleotide metabolic process Saccoglossus kowalevskii 9.76E-64 isotig ecto-like molecular_functiongo: '-nucleotidase activity Saccoglossus kowalevskii 9.76E-64 isotig ecto-like cellular_componen GO: cytoplasm Saccoglossus kowalevskii 9.76E-64 isotig ecto-like biological_process GO: cellular metabolic process Saccoglossus kowalevskii 9.76E-64 isotig ecto-like cellular_componen GO: plasma membrane Saccoglossus kowalevskii 9.76E-64 isotig ecto-like molecular_functiongo: binding Saccoglossus kowalevskii 3.16E-59 isotig ecto-like biological_process GO: nucleobase, nucleoside and nucleotide metabolic process Saccoglossus kowalevskii 3.16E-59 isotig ecto-like molecular_functiongo: '-nucleotidase activity Saccoglossus kowalevskii 3.16E-59 isotig ecto-like cellular_componen GO: cytoplasm Saccoglossus kowalevskii 3.16E-59 isotig ecto-like biological_process GO: cellular metabolic process Saccoglossus kowalevskii 3.16E-59 isotig ecto-like cellular_componen GO: plasma membrane Saccoglossus kowalevskii 3.16E-59 isotig ecto-like molecular_functiongo: binding Saccoglossus kowalevskii 9.78E-64 isotig ecto-like biological_process GO: nucleobase, nucleoside and nucleotide metabolic process Saccoglossus kowalevskii 9.78E-64 isotig ecto-like molecular_functiongo: '-nucleotidase activity Saccoglossus kowalevskii 9.78E-64 isotig ecto-like cellular_componen GO: cytoplasm Saccoglossus kowalevskii 9.78E-64 isotig ecto-like biological_process GO: cellular metabolic process Saccoglossus kowalevskii 9.78E-64 isotig ecto-like cellular_componen GO: plasma membrane Saccoglossus kowalevskii 9.78E-64 isotig ecto-like molecular_functiongo: binding Saccoglossus kowalevskii 3.17E-59 isotig ecto-like biological_process GO: nucleobase, nucleoside and nucleotide metabolic process Saccoglossus kowalevskii 3.17E-59 isotig ecto-like molecular_functiongo: '-nucleotidase activity Saccoglossus kowalevskii 3.17E-59 isotig ecto-like cellular_componen GO: cytoplasm Saccoglossus kowalevskii 3.17E-59 isotig ecto-like biological_process GO: cellular metabolic process Saccoglossus kowalevskii 3.17E-59 isotig ecto-like cellular_componen GO: plasma membrane Saccoglossus kowalevskii 3.17E-59 isotig00368 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: proton-transporting ATPase activity, rotational mechanism Pinctada fucata 0 isotig00368 atp h+ mitochondrial f1 alpha subunit cardiac muscle cellular_componen GO: mitochondrial matrix Pinctada fucata 0 isotig00368 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: ADP biosynthetic process Pinctada fucata 0 isotig00368 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: hydrogen ion transporting ATP synthase activity, rotational mechanism Pinctada fucata 0 isotig00368 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: mitochondrial ATP synthesis coupled proton transport Pinctada fucata 0 isotig00368 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: adp binding Pinctada fucata 0 isotig00368 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: mhc class I protein binding Pinctada fucata 0 isotig00368 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: embryonic development Pinctada fucata 0 isotig00368 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: plasma membrane ATP synthesis coupled proton transport Pinctada fucata 0 isotig00368 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: negative regulation of endothelial cell proliferation Pinctada fucata 0 isotig00368 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: lipid metabolic process Pinctada fucata 0 isotig00368 atp h+ mitochondrial f1 alpha subunit cardiac muscle cellular_componen GO: mitochondrial proton-transporting ATP synthase complex, catalytic core F(1) Pinctada fucata 0 isotig00368 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: eukaryotic cell surface binding Pinctada fucata 0 isotig00368 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: ATP catabolic process Pinctada fucata 0 isotig00368 atp h+ mitochondrial f1 alpha subunit cardiac muscle cellular_componen GO: plasma membrane Pinctada fucata 0 isotig00368 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: atp binding Pinctada fucata 0 isotig00369 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: proton-transporting ATPase activity, rotational mechanism Pinctada fucata 0 isotig00369 atp h+ mitochondrial f1 alpha subunit cardiac muscle cellular_componen GO: mitochondrial matrix Pinctada fucata 0 isotig00369 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: ADP biosynthetic process Pinctada fucata 0 isotig00369 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: hydrogen ion transporting ATP synthase activity, rotational mechanism Pinctada fucata 0 isotig00369 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: mitochondrial ATP synthesis coupled proton transport Pinctada fucata 0 isotig00369 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: adp binding Pinctada fucata 0 isotig00369 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: mhc class I protein binding Pinctada fucata 0 isotig00369 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: embryonic development Pinctada fucata 0 isotig00369 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: plasma membrane ATP synthesis coupled proton transport Pinctada fucata 0 isotig00369 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: negative regulation of endothelial cell proliferation Pinctada fucata 0 isotig00369 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: lipid metabolic process Pinctada fucata 0 isotig00369 atp h+ mitochondrial f1 alpha subunit cardiac muscle cellular_componen GO: mitochondrial proton-transporting ATP synthase complex, catalytic core F(1) Pinctada fucata 0 isotig00369 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: eukaryotic cell surface binding Pinctada fucata 0 isotig00369 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: ATP catabolic process Pinctada fucata 0 isotig00369 atp h+ mitochondrial f1 alpha subunit cardiac muscle cellular_componen GO: plasma membrane Pinctada fucata 0 isotig00369 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: atp binding Pinctada fucata 0 isotig00370 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: proton-transporting ATPase activity, rotational mechanism Apis mellifera 0 isotig00370 atp h+ mitochondrial f1 alpha subunit cardiac muscle cellular_componen GO: mitochondrial matrix Apis mellifera 0 isotig00370 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: ADP biosynthetic process Apis mellifera 0 isotig00370 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: hydrogen ion transporting ATP synthase activity, rotational mechanism Apis mellifera 0 isotig00370 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: mitochondrial ATP synthesis coupled proton transport Apis mellifera 0 isotig00370 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: adp binding Apis mellifera 0 isotig00370 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: mhc class I protein binding Apis mellifera 0 isotig00370 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: embryonic development Apis mellifera 0 isotig00370 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: plasma membrane ATP synthesis coupled proton transport Apis mellifera 0 isotig00370 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: negative regulation of endothelial cell proliferation Apis mellifera 0 isotig00370 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: lipid metabolic process Apis mellifera 0 isotig00370 atp h+ mitochondrial f1 alpha subunit cardiac muscle cellular_componen GO: mitochondrial proton-transporting ATP synthase complex, catalytic core F(1) Apis mellifera 0 isotig00370 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: eukaryotic cell surface binding Apis mellifera 0 isotig00370 atp h+ mitochondrial f1 alpha subunit cardiac muscle biological_process GO: ATP catabolic process Apis mellifera 0 isotig00370 atp h+ mitochondrial f1 alpha subunit cardiac muscle cellular_componen GO: plasma membrane Apis mellifera 0 isotig00370 atp h+ mitochondrial f1 alpha subunit cardiac muscle molecular_functiongo: atp binding Apis mellifera 0 isotig00371 atp synthase subunit mitochondrial precursor molecular_functiongo: proton-transporting ATPase activity, rotational mechanism Pinctada fucata 7.05E-117 isotig00371 atp synthase subunit mitochondrial precursor cellular_componen GO: mitochondrial matrix Pinctada fucata 7.05E-117 isotig00371 atp synthase subunit mitochondrial precursor biological_process GO: ADP biosynthetic process Pinctada fucata 7.05E-117 isotig00371 atp synthase subunit mitochondrial precursor molecular_functiongo: hydrogen ion transporting ATP synthase activity, rotational mechanism Pinctada fucata 7.05E-117 isotig00371 atp synthase subunit mitochondrial precursor biological_process GO: mitochondrial ATP synthesis coupled proton transport Pinctada fucata 7.05E-117