Lecture 7

Transcription
p
I

Alberts, Figure 6-9

Lecture 7

Transcription I
Lecture Outline:
1) Molecular definition
of a gene
2) Transcription and
RNA Polymerase in
prokaryotes

Readings:
Alberts,
be ts, C
Ch 6, pp
pp. 3
3299
345

Recall the flow of information in the cell

Transcription

protein synthesis
protein

Figure 6-2 Molecular Biology of the Cell ( Garland Science 2008)

Molecular Definition of a Gene

The entire nucleic acid sequence
(
(usually
ll DNA) th
thatt iis necessary ffor
the synthesis of a protein (and its
variants) or RNA
RNA. In other words
words,
genes are segments of DNA that are
transcribed into RNA
RNA. Remember
there are two types of genes that
when transcribed:
encoding
for a protein
RNA
encodes
a protein
1. RNA
functioning as RNA
functions as RNA
2. RNARNA

gene functions:
functionally,
structurally, unit of
heredity

Figure 6-3 Molecular Biology of the Cell ( Garland Science 2008)

Transcription

Alberts, Fig. 6-9

Schematic of RNA polymerase

DNA helix helix
DNA

RNA polymerase
does have 3' to 5'
exonuclease
proofreading
system

5 new RNA
5' new RNA

DNA/RNA
DNA/RNA

Alberts, Figure 6-8a

Structure of RNA polymerase

Alberts, Figure 6-8

8

Catalyzes the sequential addition of nucleotides (5 3)

base and sugar

differences
between RNA and
DNA

Base andd sugar

B
differences
between RNA and
DNA

Figure 6-4 Molecular Biology of the Cell ( Garland Science 2008)

RNA nucleotides linked by phosphodiester

bonds through ribose-PO4

1) RNA nucleotides
linked by
phosphodiester
bonds through
ribose-PO4

Generation of RNA transcript:

ssDNA
1) Template: _____
2) RNA linked by
Phosphodiester
bonds
DNA-RNA hybrid
held together by
Basepairing
ssDNA

phosphodiester
bonds

base pairing

(for bacterial
transcription)

The Transcription Cycle

Steps:
Sigma
factor binds promoter
factor
Sigma
____________
unwinding of DNA
Localized
Unwinding of DNA
localized
RNA polymerase

core RNA
polymerase

RNA polymerase
holoenzyme

sigma factor recognizes

promoter, no ATP used

Alberts, Figure 6-11

11

Transcription cycle

initial RNA
synthesized

initial RNA
synthesized

transcription does
not start at
promoter

sigma factor
released

Sigma factor
Termination
termination
released structure
structure

Steps:
RNA lsynthesis
initial
I iti
Initial
RNA synthesis
th i
factor releases
sigma
Sigma
factor releases
elongation
RNA
elongation
RNA
Termination
termination

G-C rich hairpin

loop

Alberts, Figure 6-11

12

A Question....
The RNA polymerase holoenzyme
holoenzyme......
A) consists of a sigma factor and the
polymerase
B) requires ATP to unwind the DNA
C) recognizes and binds to specific sequences
at the transcription start site
D) Answers A, B, and C are correct
E) Answers A and C are correct
13

Some key points about the transcription cycle

1) The initial steps of RNA synthesis are
inefficient
Relatively
inefficient
relatively
2) This is quite different from the elongation mode of
RNA polymerase
polymerase, which is very
very
processiveProcessive
Highly
highly
3) What are some of the characteristics of RNA
t
termination
i ti signals?
i
l ?
structure formed
hairpin
Hairpin
structure formed
rich
c following
seque
sequences
o o
g hairpin
a p
rich sequence
hairpin ces following
A-TA-T
4) How do these termination signals help to dissociate
the RNA transcript from the polymerase?
H-bonding with DNA template
disrupts
Disrupts
H-bonding with DNA template
14

Direction of transcription
Determined by the RNA polymerase
orientation of:
promoter
promoter
RNA polymerase

In this diagram, gene a is using the top DNA

strand as a template

and gene c uses the bottom DNA strand as a

template
Alberts, Fig. 6-14

15

E. coli promoters
for sigma factor
sigma^70

Pribnow box

Alberts, Figure 6-12

16

Variation in sigma subunits

17

sigma factor is in
there

Alberts, Figure 7-42a

Review Questions
How manyy sigma
g
subunits can an RNA p
polymerase
y
bind at one time?
A. 1
B. 2
C. Any number

19

Review Questions
Why is there variation in sigma subunits?
A) Different sigma units recognize and
are used by different RNA
polymerases
B) Different
Diff
t sigma
i
units
it are used
d tto
express different sets of genes
C) Different sigma units result in more
effective transcript elongation

Review Questions
What is a promoter consensus sequence?
A Th
A.
The sequence recognized
i db
by th
the RNA
holoenzyme
B. The sequence at which the RNA
transcript is initiated
C. The site on the RNA polymerase
which binds to the DNA

Simplified model of gene expression

Figure 6-21b Molecular Biology of the Cell ( Garland Science 2008)

A More Realistic Model

polycistronic

mRNA generating
multiple proteins
usually
usually product of a single promoter to
coordinate expression
Figure 6-22a Molecular Biology of the Cell ( Garland Science 2008)

Eukaryotic transcription is more complicated

Primary RNA
transcript

primary rna transcript

mature mRNA

Mature mRNA

Figure 6-21a Molecular Biology of the Cell ( Garland Science 2008)

monocystonic:
genetic info to
translate a single
protein
polycystonic:
genetic info
(mRNA) to
translate multiple
proteins, typical for
prokaryotic
sometimes
archaebacteria,
proteins have
related functions

And that is what we will talk about next class....

The End