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Gene expression and

Transcription

Learning objectives

Know the central dogma of molecular

biology.

Know the steps of transcription (in bacteria)

and describe them in general terms.

Understand why transcription a necessary

component of gene regulation.

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What is Transcription?
First step in gene expression
Segment of copied into RNA by
the RNA polymerase

RNA
Types: mRNA, rRNA and tRNA

RNA precursors

Upstream
Downstream

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RNA Structures
Primary: nuc sequence
Secondary: Base pairingds RNAs
Tertiary: ds RNAs pair again

RNA polymerase

Makes all other RNAs: Exception (**Do you remember?)

Only 1 strand of DNA is transcribed

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Promoters
RNA is transcribed only at certain DNA sites
depends on the sigma factor

Sigma binds to
both regions

RNA pol binds to s factor to recognize

promoter.

Transcription begins.

Transcription bubble formed-RNA/DNA

DS hybrid.

Termination.

Transcription

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A. Binding/Promoter recognition

1. Sigma factor + Core RNA polymerase

2. Different sigma factors can bind! (depending on
promoter)

Promoter recognition

Recognize ds DNA

-35 seq, -10 seq (AT

rich closed complex,
RP c )

Functional regions of Sigma factors

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Interaction between s70

and -35 -10

Flexible linkers binding to

aCTDs to UP elements

s3 binding to extended
-10

Isomerization

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Isomerization

-35 seq, -10 seq (AT

rich closed complex,
RP c )

RP c opens (ss DNA)

to form RP o

Initiation
Initial transcription complex

Encounters s3,2 loop, and aborts or

continues

Rifampin action

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To abort or to continue?

Elongation

Transcription elongation complex

Some hairpins form and cause backtracking

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In case of backtracking

Rnase activity
into 2ndary channel
Degrades 3 end of
RNA until clear

Termination

Types of termination
Factor independent:
1. Inverted repeat
2. Formation of hairpin
3. Hairpin is not stable

Factor dependent:
1. Terminator needed: Rho, Thau, NusA

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Factor independent termination

Transcription termination
site

Not necessarily at the end

of a gene (operon)

U-rich RNA causes RNA

Polymerase to pause,
forming hairpin loop, thus
dissociating RNA pol
from RNA

Factor dependent termination

nut termination site is not

translated

Rho binds to translated rut

Helicase activity
dissociates the RNA-DNA
binding

Formative exercise:
Identify the steps of
transcription

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Precursor of RNA

rRNA and tRNA

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