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Transcription in prokaryotes

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Page 1: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

Transcription in prokaryotes

Page 2: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA (except for the RNA primers required for replication)

Compare the mechanism of action of RNA polymerase and DNA polymerase.

[1][1]

Page 3: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

RNA POLYMERASE

’

Core Enzyme2, ’ and

Holoenzyme2, ’ and and

Page 4: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

Similarities

• Substrate for both - nucleoside triphosphates

• Pyrophosphate released for every NTP added

• Always in the 5’3’ direction

Differences

• Recognizes the site for initiation of transcription

• Ribose sugar and uracil

• No primer required

• No proofreading activity

Page 5: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

[2][2]

For genes to be transcribed by RNA polymerase the appropriate initiation site must be recognized.

I.How does the RNA polymerase know where to start transcription?

II.How does the sequence of the Pribnow box relate to its function?

Page 6: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

PROMOTERS

+1

TATAAT box

TTGACA

Transcription start site

~7 bases~19 bases

5’

3’

3’

5’

Pribnow box-35 sequence

Page 7: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

Each of the two DNA strands can serve as templates for transcription

I. What determines which of the two DNA strands will be transcribed??

II. Can both the DNA strands be transcribed??

III. Which is the sense (coding) and anti- sense (noncoding) strand ??

IV. The RNA has the same sequence as which one of the two DNA strands?

[3][3]

Page 8: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

What determines which of the two DNA strands will be transcribed??the location of the promoter

Can both the DNA strands be transcribed??Yes, but for a given gene only one strand will be the template

Which is the sense (coding) and anti- sense (noncoding) strand ??Sense strand = the same sequence as the mRNA, contains the promoterantisense = acts as the template, complementary sequence to the mRNA.

[3][3]

Page 9: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

Transcription

Page 10: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

[4][4]Three separate stages of transcription

INITIATION, ELONGATION, TERMINATION

Page 11: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required
Page 12: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required
Page 13: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

TERMINATIONindependent

Page 14: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

TERMINATION - independent

Page 15: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

TERMINATION - independent

Page 16: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

Action Of Antibiotics on Transcription

• Rifampin - binds to subunit of prokaryotic RNA polymerase– tuberculosis

• Dactinomycin – binds to DNA, prevents the movement of RNA polymerase– Tumor chemotherapy

Page 17: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

TRANSCRIPTION -TRANSCRIPTION -EUKARYOTESEUKARYOTES

Page 18: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

RNA Polymerases in Eukaryotes

RNA POLYMERASE I - rRNA

RNA POLYMERASE II - mRNA, snRNAs

RNA POLYMERASE III - tRNA and other small RNAs

Page 19: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

Eukaryotic gene Eukaryotic gene structurestructure

Exon 1 Exon 4Exon 3Exon 2 Exon 5

PROTEIN

5’ 3’

= intron (untranslated)

= exon (translated)

Pre-mRNA

Exon 1 Exon 4Exon 3Exon 2 Exon 5mRNA

Page 20: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

RNA Polymerase II

• 10-12 subunits

• The largest subunit contains a C-terminal region or domain (CTD) with 52 repeats of the amino acid sequence PTSPSYS

T = threonine

S = serine

Y = tyrosineCan be phosphorylated

Page 21: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

RNA PLOYMERSE II PROMOTERS

+1-20-30

TATA box

CAAT box

Transcription start site

25 bases45-55 bases

-70

5’

3’

3’

5’

Page 22: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

Transcription factorsTranscription factors• Any protein that regulates transcription but is not

a part of RNA polymerase, is a transcription factor

• Six general transcription factors (GTFs) required for all promoters

– TFIIA– TFIIB– TFIID– TFIIE– TFIIF– TFIIH

+ RNA Polymerase II = Preinitiation complex

Page 23: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

TATA box

TFIID BA

Pol IIF

E H

Complex of several proteins, TATA-Binding Protein (TBP) and ~12 TBP Associated Factors (TAFs)

TFIIDA TFIIA stabilizes the DNA and TFIID interactionB TFIIB binds to TFIID and acts as bridge between Pol II and TFIID

F TFIIF binds Pol II and targets its to the promoter

PREINITIATION COMPLEX (PIC)PREINITIATION COMPLEX (PIC)E Recruits TFIIHH Promoter melting, kinase activity

Page 24: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

TFIID BA

E H

Pol IIF

Page 25: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

TFIID BA

E H

Pol IIF

mRNA

GTFs only produce a basal level of transcription

Page 26: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

Enhancers/Silencers

• Upstream or downstream• Close to the promoter or thousands of base

pairs away• On either of the two strands of DNA• Act through intermediary or gene specific

transcription factors proteins• Enhancers activate transcription • Silencers deactivate transcription

Page 27: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

Enhancers/Silencers

Page 28: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

Response Elements•Enhancers contain response elements that

are responsive to certain metabolic factors

–Cyclic AMP response element (CRE)–Glucocorticoid response element (GRE)–Heat shock element (HSE)

•REs bind transcription factors produced under certain cell conditions to activate several related genes

Page 29: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

-amanitin and RNA Pol II

Page 30: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

Post Transcriptional Post Transcriptional Modification – mRNAsModification – mRNAs

5’ Capping

Page 31: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

Post Transcriptional Post Transcriptional Modification – mRNAsModification – mRNAs

Poly-A tail

Page 32: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

Post Transcriptional Post Transcriptional Modification – mRNAsModification – mRNAs

SPLICING

Page 33: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required
Page 34: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

Post Transcriptional Post Transcriptional Modification - rRNAsModification - rRNAs

RNA Polymerase I

Page 35: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

Post Transcriptional Post Transcriptional Modification - tRNAsModification - tRNAs

Page 36: Transcription in prokaryotes. In prokaryotes, only one RNA polymerase is responsible for the synthesis of all RNA ( except for the RNA primers required

THE END!


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