1. The Chapter 15 Homework is due on Wednesday, February 4 th at 11:59 pm

2. Chapter 15 Operons

3. You Must Know How the components of an operon function to regulate gene expression in both repressible and inducible operons. How positive and negative control function in gene expression.

4. Repressible and Inducible Operons: Two Types of Negative Gene Regulation 1.A repressible operon is one that is usually on; binding of a repressor to the operator shuts off transcription. 1.The trp operon is a repressible operon. Promoter trp operon Genes of operon DNA Regulatory gene mRNA Protein RNA polymerase Inactive repressor Operator Start codon Stop codon mRNA 5 trpEtrpDtrpC trpB trpA EDCBA (a) Tryptophan absent, repressor inactive, operon on Polypeptide subunits that make up enzymes for tryptophan synthesis 5 3 trpR

5. Repressible and Inducible Operons: Two Types of Negative Gene Regulation 2.An inducible operon is one that is usually off; a molecule called an inducer inactivates the repressor and turns on transcription. – Inducible enzymes usually function in catabolic pathways; their synthesis is induced by a chemical signal. – The lac operon is an inducible operon

6. E. Coli’s preferred energy source is glucose. E. Coli can use lactose as an energy source, but it has to make a series of proteins in order to do so. E. Coli will only make large amounts of the proteins necessary for lactose metabolism if the following two conditions are met. 1.Glucose levels are low 2.Lactose is available

7. The lac operon produces the proteins needed for lactose metabolism. The lacZ gene codes for  -galactosidase which hydrolyzes lactose into glucose and galactose. The lacY gene codes for the membrane protein permease which transports lactose into the cell. The lacA gene = ? IacZ IacY IacA DNA Protein mRNA 5 lac operon PermeaseTransacetylase  -Galactosidase RNA polymerase

8. Figure 15.4a DNA Regulatory gene lac I mRNA Active repressor Protein (a) Lactose absent, repressor active, operon off DNA Promoter Operator Regulatory gene No RNA made IacZ lac I mRNA RNA polymerase Active repressor Protein By itself, the lac repressor is active and switches the lac operon off.

9. Iac I Allolactose (inducer) (b) Lactose present, repressor inactive, operon on A molecule called an inducer inactivates the repressor to turn the lac operon on. IacZ IacY IacA Iac I DNA lac operon Permease Transacetylase  -Galactosidase mRNA Protein RNA polymerase mRNA 5 3 5 Inactive repressor Allolactose (inducer)

10. E. Coli’s favorite food is glucose. E. Coli will only bother to “eat” significant amounts of lactose if glucose is scarce. E. Coli will only ramp up the production of the proteins needed for lactose metabolism when glucose is scarce.

11. How E. Coli “knows” when glucose is scarce When glucose concentrations are low, cyclic AMP (cAMP), an allosteric regulatory molecule, accumulates in the cell.

12. The cAMP binds with a regulatory protein called CAP. When cAMP binds with CAP, the CAP becomes activated. Active CAP Inactive CAP cAMP

13. Figure 15.5a DNA Promoter IacZ lac I Operator RNA polymerase binds and transcribes Active CAP Inactive CAP Allolactose Inactive lac repressor CAP-binding site cAMP (a) Lactose present, glucose scarce (cAMP level high): abundant lac mRNA synthesized

14. Promoter DNA Operator IacZ lac I CAP-binding site Inactive CAP Inactive lac repressor RNA polymerase less likely to bind little lac mRNA synthesized (b) Lactose present, glucose present (cAMP level low): When glucose levels increase, CAP detaches from the lac operon, and transcription proceeds at a very low rate, even if lactose is present.