Master Biomedizin ) UCSC & UniProt 2) Homology 3) MSA 4) Phylogeny. Pablo Mier
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1 Master Biomedizin ) UCSC & UniProt 2) Homology 3) MSA 4) 1
2 12 a. All of the sequences in file1.fasta ( are homologs. How many groups of orthologs would you say there are in this file? Use Trex ( Two groups of orthologs: Protein A & protein B. b. What could you say about the history of this protein family? E.coli has only one protein, and then it duplicated to form A and B. It is possible that X.laevis_B duplicated later to form B and C. c. Would you say there is any wrongly annotated sequence? X.tropicalis_B is wrongly annotated. It should be X.tropicalis_A, because they are in the same branch. The actual X.tropicalis_B either is not in the dataset or was lost during evolution. 2
3 *Images from: Trex 12 a. Two groups of orthologs: Proteins A Proteins B b. E.coli has only one protein, and then it duplicated to form A and B. It is possible that X.laevis_B duplicated later to form B and C. c. X.tropicalis_B is wrongly annotated. It should be X.tropicalis_A, because they are in the same branch. The actual X.tropicalis_B either is not in the dataset or was lost during evolution. 3
4 13 a. Using file2.fasta ( and Trex ( can you approximate to which taxonomic division belongs proteinx? Primates. b. From which organism could it be? After guessing, check it. Homo sapiens (human) or Pan troglodytes (chimpanzee); they are 100% identical. 4
5 *Images from: Trex, UniProt 13 Drosophila melanogaster (Fruit fly) Xenopus laevis (frog) Rattus norvegicus (rat) Mus musculus (mouse) Homo sapiens (human) a. Primates Pongo abelii (orangutan) Sus scrofa (pig) Bos taurus (cattle) b. Homo sapiens (human) or Pan troglodytes (chimpanzee); they are 100% identical. 5
6 14 Human hemoglobin consists of four protein subunits: two from the alpha globin gene cluster (located on chromosome 16) and two more from the beta globin gene cluster (located on chromosome 11). But there are at least nine different globin genes in these clusters, which are: zeta, mu, alpha, theta1, epsilon, gamma1, gamma2, delta and beta. Use the proteins in file3.fasta ( a. Sort them either in cluster alpha or cluster beta. Alpha: zeta, mu, alpha, theta1. Beta: epsilon, gamma1, gamma2, delta and beta. b. Why do you think they are clustered in either cluster alpha or cluster beta? Paralogous expansion from one ancestral alpha and one ancestral beta. 6
7 *Images from: UniProt, Trex, UCSC 14 a. Using Trex: Cluster alpha Cluster beta b. Paralogous expansion from one ancestral alpha and one ancestral beta. 7
8 *Images from: Wikipedia 15 Consider the following sequences: Q9T4B2 (CYB_DELDE, dolphin), P00156 (CYB_HUMAN, human), P00157 (CYB_BOVIN, cattle), Q9MIX8 (CYB_DANRE, zebra fish), P18946 (CYB_CHICK, chicken), Q36461 (CYB_ORNAN, platypus) and P00160 (CYB_XENLA, frog). Do you think that a phylogenetic tree built using these sequences would be coherent with evolution? Check with the taxonomy provided by UniProt ( Human (Homo sapiens) Dolphin (Delphinus delphis) Cattle (Bos taurus) Chicken (rooster) (Gallus gallus) 8 Zebra fish (Danio rerio) Platypus (Ornithorhynchus anatinus) Frog (Xenopus laevis)
9 *Images from: UniProt, Trex 15 Dolphin Eukaryota Metazoa Chordata Craniata Vertebrata Euteleostomi Mammalia Eutheria Laurasiatheria Cetartiodactyla Cetacea Odontoceti Delphinidae Delphinus Cattle Eukaryota Metazoa Chordata Craniata Vertebrata Euteleostomi Mammalia Eutheria Laurasiatheria Cetartiodactyla Ruminantia Pecora Bovidae Bovinae Bos Human Eukaryota Metazoa Chordata Craniata Vertebrata Euteleostomi Mammalia Eutheria Euarchontoglires Primates Haplorrhini Catarrhini Hominidae Homo Platypus Eukaryota Metazoa Chordata Craniata Vertebrata Euteleostomi Mammalia Monotremata Ornithorhynchidae Ornithorhynchus Frog Eukaryota Metazoa Chordata Craniata Vertebrata Euteleostomi Amphibia Batrachia Anura Pipoidea Pipidae Xenopodinae Xenopus Xenopus Chicken Eukaryota Metazoa Chordata Craniata Vertebrata Euteleostomi Archelosauria Archosauria Dinosauria Saurischia Theropoda Coelurosauria Aves > Gallus Zebrafish Eukaryota Metazoa Chordata Craniata Vertebrata Euteleostomi Actinopterygii Neopterygii Teleostei Ostariophysi Cypriniformes Cyprinidae Danio 9
10 MSA + + Homology 16 Consider the following sequences: P50225, P50226, P0DMM9, P0DMN0, O43704, O00338, Q6IMI6, O All of them are part of a human paralog family (sulfotransferases). a. Look for the coordinates of the substrate binding region in the UniProt entry P Do you expect all the paralogs to share this substrate binding region? Why? Check it. Yes, because they share the motif K[ST]H. b. Using the previous sequences, could you determine which human paralog is the ortholog of the sequence in file4.fasta ( Do not use BLAST. The ortholog of protein_of_interest is ST1B1_HUMAN. c. Which protein is the one found in file4.fasta? ST1B1_CANLF, from Canis familiaris (dog). 10
![the motif K[ST]H. b.](/docs-images/87/95056720/images/11-6.jpg "The ortholog of")
11 MSA + + Homology 16 *Images from: UniProt, Trex (P50225) a. Yes, because they share the motif K[ST]H. b. The ortholog of protein_of_interest is ST1B1_HUMAN. c. ST1B1_CANLF, from Canis familiaris (dog). 11
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