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Lecture 7
Transcription I
Alberts, Figure 6-9 1

Lecture 7
Transcription I
Lecture Outline: Readings:

g
MBoC, Ch 6 - pp. 329-
1)) Molecular definition 345
of a gene
2)) Transcription
p and
RNA Polymerase in
y
prokaryotes
Recall the flow of information in the cell
Transcription
Figure 6-2 Molecular Biology of the Cell (© Garland Science 2008)

Molecular Definition of a Gene
The entire nucleic acid sequence
(usually DNA) that is necessary for
the synthesis of a protein (and its
variants) or RNA. In other words,
genes are segments of DNA that are
transcribed into RNA. Remember
there are two types of genes that
when transcribed:
The resulting
1. Resulting RNA encodesRNA
a proteinencodes a protein
The
2. End resulting
product is an RNA RNA functions as RNA
A has more promoters for it.

Transcription
a control method that modifies methods of expression
more promoters means more proteins and more frequently transcribed

product of this transcription is ribosomal RNA
polymerase is moving from left to right
Alberts, Fig. 6-9 6

Schematic of RNA polymerase
DNA helix
(NTPs)
5’ new RNA DNA/RNA

Alberts, Figure 6-8a
7
Structure of RNA polymerase
Alberts Figure 6
Alberts, 6-8
8
8
Catalyzes the sequential addition of nucleotides (5’ – 3’)
Base and sugar

ddifferences
e e ces
1) RNA nucleotides
l tid
between RNA and
linked by
DNA
phosphodiester
h h di t
bonds through
ribose-PO
ib PO4

Generation of RNA transcript:
1) Template: _____ ssDNA

3'-5' single-
stranded DNA
2) RNA linked by
 phosophodiester
Phosphodiester bonds
bonds
• DNA-RNA hybrid
held together
g byy
 H-bonding
Basepairing
through base pairs
The Transcription Cycle
Steps:
 sigma
Sigma
____________factor
binds to DNA binds promoter
 shows
Locali
Localized ed Unwinding
Un where
RNA polymerase inding
to startof DNA
transcription,
positions it in front of different genes
RNA polymerase

Transcription cycle
initial RNA
Holoenzyme =
RNAP + sigma
synthesized
Sigma factor
C-Gs form H-bonds
Termination
released
structure
Steps: assists in positioning by binding to promotors
 Initial
Initial
RNARNA synthesis
y Transcription start, sigma factor dissociates, abortive
synthesis;
translation
 Sigma
Sigma factorfactor releases
is released
 RNA
RNA elongation
polymerase g reaches a stop codon
 Usually
Termination
has G-C rich regions and then A-T rich regions
Some key
yppoints about the transcription
p cycle
y
1) The initial steps of RNA synthesis are are…

 sigma
Relatively inefficient
factor releases , elongation, termination (relatively efficient)
2) This is quite different from the elongation mode of
RNA polymerase, which is very…
 highly
Highly Processive
proccessive - very fast
3) What are some of the characteristics of RNA

termination signals?
 Rho
Hairpin structure
independent terminationformed
- formation of hairpin structures, A-T rich within
 Dependent:
A-T richdisrupts
sequences
H-bonding following hairpin
with DNA template between RNAP at the active
H sitedo
4) How d these
th termination
t i ti signals i l help
h l to t dissociate
di i t
the RNA transcript from the polymerase?
 breaks
Disrupts
H-bondsH-bonding with template
on active site between DNA template
and polymerase
13
Direction of transcription
The transcript
generated usingof gene
bottom
Determined by the  asRNA
3' - 5' polymerase
direction,
a template
strand you use
f
strand as template
and gene g use the
orientation of: bottom DNA strand
 position
promoterof RNAP to which strand
orientation of the promoter as the
th template
t l t
- usually asymetric
- only one possible
Thetop
Uses transcript
strand as aof gene
a is
template
generated using the top DNA
strand as a template Alberts, Fig. 6-14 14
E coli promoters
E.
+1 is the front end of an RNA being made. upstream of this position are -35 and -10
promoters - they are hexomeric (have six nucleotides)
Alberts Figure 6
Alberts, 6-12
12
15
Variation in sigma subunits
16
Alberts, Figure 7-42a
Simplified
p model of ggene expression
p
Figure 6-21b Molecular Biology of the Cell (© Garland Science 2008)

A More Realistic Model
• polycistronicmRNA generating
multiple proteins
•usually
y product
p of a single
g promoter
p to
coordinate expression
Figure 6-22a Molecular Biology of the Cell (© Garland Science 2008)
Eukaryotic
y transcription
p is more complicated
p
Primary RNA
transcript
Mature mRNA
Figure 6-21a Molecular Biology of the Cell (© Garland Science 2008)

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

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Lecture 7

Alberts, Figure 6-9 1

Lecture Outline: Readings:

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

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

polymerase is moving from left to right

Alberts, Fig. 6-9 6

5’ new RNA DNA/RNA

Base and sugar

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

1) Template: _____ ssDNA

Alberts, Figure 6-11 11

1) The initial steps of RNA synthesis are are…

3) What are some of the characteristics of RNA

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

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

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