1. Sites of regulation

2. Feedback inhibition

3. Mechanism of allosteric inhibition

4. Repression and Induction

5. Mechanism of repression- Negative control

6. Mechanism of repression- control by co-repressor Operon= a cluster of genes under control of a single promoter Regulon? Arginine synthesis

7. Mechanism of induction Lac operon

8. Mechanism of induction- negative control

9. Positive vs. Negative control Repressors are “negative control” –An active repressor (- inducer or + corepressor) stops transcription Activator proteins are “positive control” –The regulatory protein (activator) promotes transcription –Example: maltose regulon

10. Activator protein without inducer- positive control

11. Activator protein with inducer

12. DNA binding proteins Non-specific, eg. histones –Small proteins, high + charge Specific –Frequently dimers –Interact with inverted repeats –Eg. lac repressor

13. DNA binding proteins Dimeric proteins (e.g., lac repressor) interact with inverted repeats

15. Attenuation Positive and negative control affect initiation of transcription Attenuation affects continuation of transcription –Eg. the tryptophan operon has a leader that includes two tryptophan residues –When tryptophan is lacking, the translation is delayed –The speed of translation determines which of two mRNA double-stranded loops form One of the two possible loops is a termination signal

16. How does it work? Transcription and translation occurring almost simultaneously –Rate of transcription influenced by rate of translation Translation of leader PEPTIDE regulates transcription –Synthesis of leader terminates transcription, and leader synthesis is inhibited by low Trp

17. Attenuation: leader sequence

18. Attenuation: delayed translation Ribosome pauses at trp codon, stem loop that forms DOES not terminate transcription

19. Attenuation: undelayed translation Leader peptide is formed Stop codon or stem-loop structure can form in mRNA And transcription is attentuated

20. Global control: catabolite repression- a variety of unrelated genes regulated Diauxic growth

21. Catabolite repression Catabolite activator protein (CAP) assists binding of RNA polymerase to promoter CAP can bind only when it first binds cAMP Adenylate cyclase: ATP -> cAMP + pyrophosphate Glucose inhibits adenylate cyclase and stimulates cAMP excretion Catabolite repression is similar to positive control, but the difference is the global nature of catabolite repression

22. CAP binding site on the lac operon

23. Quorum sensing Also a form of global control Relatively recent discovery AHL-acylated homoserine lactone –Diffusible –Inducer needs activator protein Example, bioluminescence and luxR activator –Only when [AHL ] is high enough will LuxR activate the lux operon

24. 2 component regulatory systems Maltose=effector, BUT if signal not DIRECTLY involved, but needs to be transmitted and changed = signal transduction Sensor protein= –kinase, phosphorylates compounds, –membrane associated Phosphoryl group transmitted to another regulator IN the cell –Often a DNA binding protein involved in transcription Many examples, N-fixation, sporulation,chemotaxis

25. 2 component regulatory systems

26. Chemotaxis Attractants decrease rate of autophosphorylation Repellant increased autophosphorylation CheA-CheW=transducer CheY controls switch –cheY-P tumbles, CCW-CW CheB phosporylated by CheA-P, but slower response than CheY-P CheB involved methylation –Fully methylated = best for repellants –cheB-P demethylates, occurs when attractants High –Degree of methylation regulates attraction/repulsion

27. Chemotaxis