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15
Regulation of
Gene Expression
Lecture Presentations by
Kathleen Fitzpatrick and Nicole Tunbridge
Enzyme 1 Regulation
trpD gene of gene
expression
trpB gene
Enzyme 3
trpA gene
Tryptophan
DNA
No RNA
made
mRNA
Protein Active
repressor
Tryptophan
(corepressor)
(b) Tryptophan present, repressor active, operon off
© 2014 Pearson Education, Inc.
Figure 15.3a
DNA
No RNA
made
mRNA
Protein Active
repressor
Tryptophan
(corepressor)
(b) Tryptophan present, repressor active, operon off
Active
Protein
repressor
(a) Lactose absent, repressor active, operon off
lac operon
RNA polymerase
3
mRNA mRNA 5
5
Allolactose Inactive
(inducer) repressor
(b) Lactose present, repressor inactive, operon on
© 2014 Pearson Education, Inc.
Figure 15.4a
Regulatory Promoter
gene Operator
DNA lacI IacZ
No
RNA
3 made
mRNA RNA
5 polymerase
Active
Protein
repressor
(a) Lactose absent, repressor active, operon off
RNA polymerase
mRNA 3
mRNA 5
5
Protein
-Galactosidase Permease Transacetylase
Inactive
repressor
Allolactose
(inducer)
(b) Lactose present, repressor inactive, operon on
Inactive lac
Inactive
repressor
CAP
Allolactose
(a) Lactose present, glucose scarce (cAMP level high):
abundant lac mRNA synthesized
Promoter
Promoter
Inactive lac
Inactive
repressor
CAP
Allolactose
(a) Lactose present, glucose scarce (cAMP level high):
abundant lac mRNA synthesized
Promoter
Inactive
CAP Inactive lac
repressor
NUCLEUS
Chromatin
Chromatin modification:
DNA unpacking involving
histone acetylation and
DNA demethylation
DNA
Gene available
for transcription
Gene
Transcription
RNA Exon
Primary transcript
Intron
RNA processing
Tail
Cap
mRNA in nucleus
Transport to cytoplasm
CYTOPLASM
mRNA in cytoplasm
Degradation Translation
of mRNA
Polypeptide
Protein processing, such
as cleavage and
chemical modification
NUCLEUS
Chromatin
Chromatin modification:
DNA unpacking involving
histone acetylation and
DNA demethylation
DNA
Gene available
for transcription
Gene
Transcription
RNA Exon
Primary transcript
Intron
RNA processing
Tail
Cap mRNA in nucleus
Transport to cytoplasm
CYTOPLASM
© 2014 Pearson Education, Inc.
Figure 15.6b
CYTOPLASM
mRNA in cytoplasm
Degradation Translation
of mRNA
Polypeptide
Protein processing, such
as cleavage and
chemical modification
Nucleosome
Histone
tails
Coding segment
mRNA G P P P AAA AAA 3
Start Stop
5 Cap 5 UTR codon codon 3 UTR Poly-A
tail
DNA
Upstream
Promoter
Transcription
termination
region
Downstream
© 2014 Pearson Education, Inc.
Figure 15.8b-1
Proximal Poly-A signal
control Transcription sequence
elements start site
Exon Intron Exon Intron Exon
DNA
Promoter
DNA
Promoter
Transcription
Poly-A
signal
Primary RNA Exon Intron Exon Intron Exon
transcript 5
(pre-mRNA) Cleaved 3
end of
primary
transcript
DNA
Promoter
Transcription
Poly-A
signal
Primary RNA Exon Intron Exon Intron Exon
transcript 5
(pre-mRNA) Cleaved 3
RNA end of
processing primary
Intron RNA
transcript
Coding segment
mRNA G P P P AAAAAA 3
Start Stop
5 Cap 5 UTR codon codon 3 UTR Poly-A
tail
© 2014 Pearson Education, Inc.
The Roles of Transcription Factors
Activation
domain DNA-binding
domain
DNA
Chromatin modification
Transcription
RNA processing
mRNA Translation
degradation
Protein
processing
and degradation
Activators Promoter
Gene
DNA
Distal control
Enhancer TATA box
element
Activators Promoter
Gene
DNA
Distal control
Enhancer TATA box
element
General transcription
factors
DNA-
bending
protein
Group of mediator proteins
Activators Promoter
Gene
DNA
Distal control
Enhancer TATA box
element
General transcription
factors
DNA-
bending
protein
Group of mediator proteins
RNA
polymerase II
RNA
polymerase II
Transcription
initiation complex RNA synthesis
© 2014 Pearson Education, Inc.
Some transcription factors function as repressors,
inhibiting expression of a particular gene by a
variety of methods
Some activators and repressors act indirectly by
influencing chromatin structure to promote or
silence transcription
Control
elements
Enhancer Promoter Crystallin gene
Albumin gene
Albumin gene not expressed
expressed
Crystallin gene
not expressed Crystallin gene
expressed
© 2014 Pearson Education, Inc.
Figure 15.11a
Albumin gene
expressed
Crystallin gene
not expressed
© 2014 Pearson Education, Inc.
Figure 15.11b
(b) LENS CELL NUCLEUS
Available
activators
Albumin gene
not expressed
Crystallin gene
expressed
© 2014 Pearson Education, Inc.
Coordinately Controlled Genes in Eukaryotes
Chromatin modification
Transcription
RNA processing
mRNA Translation
degradation
Protein
processing
and degradation
Exons
DNA 1 2 3 4 5
Troponin T gene
Primary
RNA 1 2 3 4 5
transcript
RNA splicing
mRNA 1 2 3 5 or 1 2 4 5
© 2014 Pearson Education, Inc.
mRNA Degradation
Chromatin modification
Transcription
RNA processing
mRNA Translation
degradation
Protein
processing
and degradation
50 m
DNA in nucleus
1 Test tube containing
reverse transcriptase
and mRNA mRNAs in
cytoplasm
DNA in nucleus
1 Test tube containing
reverse transcriptase
and mRNA mRNAs in
cytoplasm
Reverse
transcriptase Poly-A tail
2 Reverse transcriptase mRNA
makes the first 5 A A A A A A 3
3 T T T T T 5
DNA strand.
DNA Primer
strand
DNA in nucleus
1 Test tube containing
reverse transcriptase
and mRNA mRNAs in
cytoplasm
Reverse
transcriptase Poly-A tail
2 Reverse transcriptase mRNA
makes the first 5 A A A A A A 3
3 T T T T T 5
DNA strand.
DNA Primer
strand
3 mRMA is degraded.
5 A A A A A A 3
3 T T T T T 5
DNA in nucleus
1 Test tube containing
reverse transcriptase
and mRNA mRNAs in
cytoplasm
Reverse
transcriptase Poly-A tail
2 Reverse transcriptase mRNA
makes the first 5 A A A A A A 3
3 T T T T T 5
DNA strand.
DNA Primer
strand
3 mRMA is degraded.
5 A A A A A A 3
3 T T T T T 5
4 DNA polymerase
synthesizes the
5 3
second strand. 3 5
DNA
polymerase
DNA in nucleus
1 Test tube containing
reverse transcriptase
and mRNA mRNAs in
cytoplasm
Reverse
transcriptase Poly-A tail
2 Reverse transcriptase mRNA
makes the first 5 A A A A A A 3
3 T T T T T 5
DNA strand.
DNA Primer
strand
3 mRMA is degraded.
5 A A A A A A 3
3 T T T T T 5
4 DNA polymerase
synthesizes the
5 3
second strand. 3 5
DNA
polymerase
5 cDNA carries complete
5 3
coding sequence 3 5
without introns. cDNA
© 2014 Pearson Education, Inc.
Figure 15.16
Technique
1 cDNA synthesis mRNAs
cDNAs
Primers
2 PCR amplification
-globin
gene
3 Gel electrophoresis
Embryonic stages
Results 1 2 3 4 5 6
Genes in red
wells expressed
in first tissue.
Genes in green
wells expressed
in second tissue.
Genes in yellow
wells expressed
in both tissues.
Genes in black
wells not
DNA microarray expressed in
either tissue.
Transcription
• Alternative RNA splicing:
Primary RNA
RNA processing transcript
mRNA or
mRNA Translation
degradation
Translation
Protein processing
and degradation • Initiation of translation can be controlled
via regulation of initiation factors.
Chromatin modification
Transcription
RNA processing
mRNA Translation
degradation
Protein processing
and degradation
Transcription
• Regulation of transcription initiation:
DNA control elements in enhancers bind
specific transcription factors.
Bending of
the DNA
enables
activators to
contact proteins at
the promoter, initiating transcription.
• The genes in a coordinately controlled
group all share a combination of control
elements.
Enhancer Promoter
Gene 1
Gene 2
Gene 3
Gene 4
Gene 5