32 Gene regulation, continued Lecture Outline 11/21/05
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1 32 Gene regulation, continued Lecture Outline 11/21/05 Review the operon concept Repressible operons (e.g. trp) Inducible operons (e.g. lac) Positive regulation of lac () Practice applying the operon concept to predict: the phenotypes of mutants The characteristics of other operons Gene regulation in prokaryotes vs eukaryotes The trp operon: Tryptophan absent -> inactive -> transcription Trp operon Active can bind to operator and block transcription Regulatory gene 5 trpr Figure 18.21a 3 RNA polymerase The operator is a particular sequence of bases where the can bind Promoter Operator 5 trp operon Genes of operon trpe trpd trpc trpb trpa E D C B A One long codes several Polypeptides that make up polypeptides, each with its own start enzymes for tryptophan synthesis and stop codon Figure 18.21b Tryptophan (co) Active No RNA made Tryptophan present -> active -> operon off. 1
2 Lac operon Inducible operons are normally off When lactose is present, can no longer bind. Transcription occurs Positive vs Negative Gene Regulation Both the trp and lac operons involve negative control of genes because the operons are switched off by the active form of the protein Some operons are also subject to positive control An activator protein is required to start transcription. E.g. catabolite activator protein () Positive Gene Regulation- In E. coli, glucose is always the preferred food source When glucose is scarce, the lac operon is activated by the binding of Active form of helps RNA polymerase bind to promoter, so transcription can start lacl camp Inactive Promoter Active lacz Operator Inactive lac Figure 18.23a G-protein-linked receptor GTP You ve seen camp used in other signaling pathways First messenger (signal molecule such as epinephrine) G protein ATP camp Adenylyl cyclase Cellular responses Enzyme adenylyl cyclase kinase A 2
3 When glucose is abundant, camp is used up detaches from the lac operon, prevents RNA polymerase from binding to the promoter Glucose transporter complex also activates adenylate cyclase Promoter lacl lacz Operator RNA polymerase can t bind Inactive Figure 18.23b Inactive lac If it is busy phosphorylating glucose, it cannot activate adenylate cyclase, so level of camp falls How do genetic switches work? binding proteins can be either s or activators, depending on how they intereact with RNA polymerase Activator This configuration helps RNA polymerase bind Repressor This configuration blocks RNA polymerase 3
4 Dual control of the lac operon Glucose must be absent Lactose must be present + glucose + lactose + glucose - lactose - glucose - lactose - glucose + lactose off, because not bound off, because active and not bound off, because active Operon active The Lac operon X-ray structure of camp bound to 5 lacl lacz lacy laca 3 RNA polymerase 5' 5 Many Operons use lac, gal, mal, ara, etc. Allolactose (inducer) Inactive β-galactosidase Permease Transacetylase binds to RNA polymerase Figure 18.22b What will happen if there is a deletion of the: + lactose? - lactose? operator? lac gene? binding site? 4
5 Arabinose is another sugar that E. coli can metabolize Will those genes be repressible or inducible? How might it be regulated? Arabinose can bind to the Arginine is an essential amino acid. Will that pathway be repressible or inducible? How might argenine synthesis be regulated? O Galactose is yet another sugar that E. coli can metabolize. Will those genes be repressible or inducible? How might gal be regulated? P Galactose O gale galt galk Epimerase Transferase Kinase Gal protein (galr) Don t memorize these names- just the general concept. Gene Regulation in Prokaryotes and Eukarykotes Prokaryotes Operons 27% of E. coli genes (Housekeeping genes not in operons) simultaneous transcription and translation Eukaryotes No operons, but they still need to coordinate regulation More kinds of control elements RNA processing Chromatin remodeling Histones must be modified to loosen 5
6 Signal NUCLEUS Chromatin Packing Cap Chromatin modification: Gene Transcription RNA Exon Degradation of Intron RNA processing Tail Transport to cytoplasm CYTOPLASM in cytoplasm Translation Gene available for transcription Primary transcript in nucleus Nucleosome (b) 30-nm fiber scaffold (c) Looped domains (300-nm fiber) 30 nm 300 nm Loops Scaffold Polypetide Cleavage Chemical modification Transport to cellular destination Active protein 700 nm Degradation of protein Figure 19.3 Degraded protein 1,400 nm Figure 19.2 Histone Modification Chromatin changes Histone acetylation loosens to allow transcription Transcription RNA processing Translation degradation processing and degradation Histone tails Figure 19.4a double helix Amino acids available for chemical modification Figure 19.4 b Unacetylated histones Acetylated histones 6
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