Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643...

26
Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 [email protected]

Transcript of Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643...

Page 1: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

Prokaryotic Gene Regulation

April 22, 2015

Richard D. Howells, PhD

MSB E-643

[email protected]

Page 2: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

Objectives

During this presentation, you may catch up on needed sleep recall the process of transcription appreciate how the lac and trp operons are

regulated in bacteria

Lecture 13 2

Page 3: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

3

General structure of the tandem arrays of pre-rRNA transcription units

Page 4: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

44

RNA is synthesized 5’ to 3’ based on the DNA sequence of the template DNA strand

Page 5: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

5

The bacterial RNA polymerase

Page 6: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

6

Bacterial RNA Polymerase

Page 7: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

7

Inactivation of theprokaryotic RNA

polymerase byrifampin - a

drug used for treatment of tuberculosis

Page 8: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

8

Prokaryotic promoter region: landmark for site of transcription initiation

Page 9: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

9

The Lactose (Lac) Operon of Escherichia coli

Lac I encodes lac repressor

Lac Z encodes b-galactosidase

Lac Y encodes lac permease

Lac A encodes b-galactoside transacetylase

1920-2013

Page 10: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

10

Lactose metabolism

in E. coli

Page 11: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

11

Transcriptional Regulation of the Lac Operon

Page 12: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

18

The Lac Operon Promoter

Page 13: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

The lac promoter vs. the promoter consensus sequence

13

Page 14: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

14

Structure of cAMP

Page 15: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

15

Control of the lac operon by glucose and cAMP

Page 16: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

cAMP-CAP (CRP) homodimer (yellow domain interacts with RNA pol)

16

Page 17: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

tTThe T

17

The cAMP-CAP complex activates transcription

of the lac operon

Page 18: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

18

Lac Operon Regulation

Page 19: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

Regulation of the trp operon in E. coli

19

Page 20: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

Trp repressor homodimer bound to Trp and the trp operator

20

Page 21: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

21

Rho-independenttermination oftranscription

Page 22: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

Transcriptional attenuation in the trp operon

22

Page 23: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

Transcriptional attenuation in the trp operon

23

Page 24: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

Transcriptional attenuation in the trp operon

24

Page 25: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

Attenuation in the trp operon

25(does not cause attenuation)

Page 26: Prokaryotic Gene Regulation April 22, 2015 Richard D. Howells, PhD MSB E-643 howells@njms.rutgers.edu.

Leader sequences of other AA operons in E. coli subject to attenuation

26