UNIT 6 PART 3 *REGULATION USING OPERONS* Hillis Textbook, CH 11

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1 UNIT 6 PART 3 *REGULATION USING OPERONS* Hillis Textbook, CH 11

2 REVIEW: Signals that Start and Stop Transcription and Translation BUT, HOW DO CELLS CONTROL WHICH GENES ARE EXPRESSED AND WHEN? First of all, There is a difference between regulation in a prokaryote and a eukaryote.

3 OPERONS - PROKARYOTES Prokaryotes conserve energy by making proteins only when needed. The most efficient gene regulation is at the level of transcription. A gene cluster with a single promoter is an operon. An operator is a short stretch of DNA near the promoter that controls transcription of the structural genes. 1. Inducible operon turned off unless needed In inducible systems a metabolic substrate (inducer) interacts with a regulatory protein (repressor); the repressor cannot bind and allows transcription. Usually control CATABOLIC REACTIONS 2. Repressible operon turned on unless not needed In repressible systems a metabolic product (co-repressor) binds to regulatory protein, which then binds to the operator and blocks transcription. Usually control ANABOLIC REACTIONS.

4 LAC OPERON - INDUCIBLE A compound that induces protein synthesis is an inducer. When the enzymes are induced, metabolism will take place.

5 LAC OPERON INDUCIBLE E. coli must adapt quickly to supply of food (lactose is a dissacharide example) Uptake and metabolism of lactose involves three important -galactoside enzymes -galactoside is a type of glycosidic bond between monosaccharides if this is present, LACTOSE is present. If E. coli is grown with glucose but no lactose present, no enzymes for lactose conversion are produced. If lactose is predominant and glucose is low, E. coli synthesizes all three enzymes. If lactose is removed, synthesis stops.

6 LAC OPERON INDUCIBLE The lac operon is only transcribed when a - galactoside predominates in the cell: A repressor protein is normally bound to the operator, which blocks transcription. In the presence of a -galactoside, the repressor detaches and allows RNA polymerase to initiate transcription. The key to this regulatory system is the repressor protein.

7 NO lactose, repressor will not allow transcription = NO enzymes to metabolize lactose. WITH lactose available, the repressor is removed, transcription will allow expression of enzymes to metabolize lactose

8 TRP OPERON REPRESSIBLE A repressible operon is switched off when its repressor is bound to its operator. However, the repressor only binds in the presence of a co-repressor. The co-repressor causes the repressor to change shape in order to bind to the promoter and inhibit transcription. Tryptophan functions as its own corepressor, binding to the repressor of the trp operon.

9 NO trp = no repressor and transcription takes place, allowing enzymes to be synthesized for tryptophan. WITH trp, the repressor becomes activated and no transcription takes place.

10 UNIT 6 PART 3 *REGULATION USING TRANSCRIPTON FACTORS* Hillis Textbook, CH 11

11 TRANSCRIPTION FACTORS - EUKARYOTES Genes can be regulated at the level of transcription. Two types of regulatory proteins, called transcription factors, control whether a gene is active. These proteins bind to specific DNA sequences near the promoter: 1. Negative regulation prevents transcription 2. Positive regulation stimulates transcription THIS IS HOW EUKARYOTIC GENES ARE TURNED ON AND OFF!

12 An activator protein binds to stimulate transcription A repressor protein prevents transcription

13 TRANSCRIPTION FACTORS: Transcription factors act at eukaryotic promoters. Each promoter contains a core promoter sequence where RNA polymerase binds. TATA box is a common core promoter sequence rich in A-T base pairs. Only after general transcription factors bind to the core promoter, can RNA polymerase II bind and initiate transcription.

14 REGULATORY PROTEINS: Besides the promoter, other DNA sequences can bind regulatory proteins that interact with RNA polymerase and regulate transcription. Some are positive regulators activators (DNA sequence is called an enhancer); others are negative repressors (DNA sequence is called a silencer).

15 ALTERNATIVE SPLICING: Eukaryotic gene expression can be regulated after the initial gene transcript is made. Different mrnas can be made from the same gene by alternative splicing (as introns and exons are spliced out, new proteins are made). Mechanism for generating proteins with different functions, from a single gene.

16 TRANSLATIONAL REGULATION: Three ways to regulate mrna translation: Inhibition of translation with mirnas Modification of the 5 cap end of mrna can be modified if cap is unmodified mrna is not translated. Repressor proteins can block translation directly translational repressors

17 Posttranslational aspects of protein synthesis: Polypeptide emerges from the ribosome and folds into its 3-D shape. Its conformation allows it to interact with other molecules it may contain a signal sequence indicating where in the cell it belongs (nucleus, mitochondria, etc.) In the absence of a signal, the protein will remain where it was produced.

18 Protein modifications: Proteolysis cutting of a long polypeptide chain into final products, by proteases. Glycosylation addition of carbohydrates to form glycoproteins Phosphorylation addition of phosphate groups catalyzed by protein kinases charged phosphate groups change the conformation of the protein

19 LAC OPERON - INDUCIBLE

20 LAC OPERON - INDUCIBLE

21 TRP OPERON - REPRESSIBLE

22 TRP OPERON - REPRESSIBLE

23 TRANSCRIPTION FACTORS:

24 TRANSCRIPTION FACTORS:

25 TRANSCRIPTION FACTORS (TATA BOX):

26 REGULATORY PROTEINS:

27 ALTERNATIVE SPLICING:

28 TRANSLATIONAL REGULATION:

29 PROTEIN MODIFICATIONS:

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