Regulation of Gene

Expression

AP Biology
Ch 15
 Gene  Protein Control
• Feedback inhibition –
enough product is made
the system shuts down
– More product is made
when needed
– The product shuts down
the process
• Gene Expression –
genes are only
expressed when needed.
Often regulated at
transcription.
 Gene Expression: Prokaryotes
• Operon – grouped genes that are transcribed together –
code for functionally similar proteins
• Key Players
– Promoter – section of DNA where RNA polymerase binds
– Operator – Controls activation of transcription
• on off switch
• between promoter and genes for proteins – structural genes
– Repressor protein – binds to operator to block RNA polymerase
and shut down transcription
• Turns off the operon
• Corepressor – keeps the repressor protein on the operator
– Trp operon
• Inducer – pulls repressor off the operator
– Turns on the operon – lactose on the lac operon
– Regulatory gene – produces the repressor protein
– Structural genes – code for proteins
Positive and Negative Gene Regulation
• Negative • Positive
– Repressible: usually on but – E. coli prefer to use glucose for
can be inhibited trp operon, energy, they will only use lactose
allosteric inhibition, when glucose is in short supply
tryptophan present prevents – glucose cAMP binds to
its own production. regulatory protein “CAP” & stimulates
(anabolic) gene transcription
– Inducible: usually off, but Positive gene regulation!
can be turned on, an inducer – The cAMP & CAP combination allow
(a specific small molecule, RNA polymerase to bind to the
allolactose in the lac operon) promoter sequence more efficiently.
inactivates the repressor and – Remember cAMP is regulating the
allows transcription gene expression in the bacteria
(catabolic)
Trp operon: repressible,
always making tryptophan,
repressed if tryptophan is
“eaten” tryptophan is
necessary for the cell to
function
Lac operon: inducible,
only turned on if lactose is
“eaten” lactose is not
necessary for the cell to
function
 Eukaryotic Chromosome
• Chromosomes – tightly coiled DNA
around proteins during cell division
• Chromatin – loosely packed DNA
around proteins
• Histones – protein which the DNA
wraps around
• Nucleosomes – grouped histones
together
– Heterochromatin – tighter packed
chromatin
• Not transcribing
– Euchromatin – looser packed chromatin
• Transcription occurring
Gene Expression: Eukaryotes
• Cell Differentiation –
cell specialization
• All cells contain the
same genes
• The genes that are
expressed determines
the type of cell
– Ex: Skin cell vs. a nerve
cell
Chromatin Regulation
• Histone acetylation –
allows transcription factors
to bind to DNA allowing
transcription to occur
– Creates loosely packed DNA
- euchromatin
• DNA Methylation – occurs
after DNA synthesis has
occurred
– Lower transcription rates
– One X in females is highly
methylated
– Works w/ a deacetylation
enzyme in some spp.
 Epigenetic inheritance
• Not controlled by base
sequences.
• DNA methylation
(deactivates one
homologous chromosome)
may explain abnormal or
unexpected DNA
expression as is often seen
in identical twins.
http://images.the-scientist.com/content/images/general/55342-1.jpg
 Regulation of Transcription

• Transcription involves RNA Polymerase II and

transcription factors
• RNA polymerase II attaches to the promoter
(TATA box) sequence to begin transcription
• Control elements – non coding sequences of
DNA where the transcription factors attach
 Regulation of Transcription
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• Enhancer – control element far from a

gene or intron
• Activator – bind to enhancers to turn on
transcription of a gene
• Transcription factors + enhancer + activator
+ RNA Polymerase II = transcription
initiation complex
– Needed for transcription to begin
• Repressors – inhibit gene expression
– Turn off transcription
– Block activators from binding to enhancers
 Distal control
element
Promoter
Activators
Gene

Enhancer TATA
box
1 General
Activator proteins bind transcription
to distal control elements factors
grouped as an enhancer in
the DNA. This enhancer has
three binding sites. DNA-bending
protein

2 Group of
A DNA-bending protein Mediator proteins
brings the bound activators
closer to the promoter.
Other transcription factors,
mediator proteins, and RNA RNA
polymerase are nearby. Polymerase II

Chromatin changes

3
The activators bind to Transcription

certain general transcription RNA processing

factors and mediator RNA

proteins, helping them form Polymerase II
an active transcription mRNA
degradation
Translation

initiation complex on the promoter. Protein processing

and degradation

Transcription RNA synthesis

Initiation complex
 RNA Processing Regulation
• Alternative RNA Splicing – different regions of the pre-
mRNA serve as introns or exons creating different mRNA
strands depending on what is removed & spliced together.
 mRNA Degredation
• Prokaryotes • Eukaryotes
– Short Life span – Survive from hours to
– Degraded in seconds weeks
– Allows rapid response to – Internal conditions
environmental changes constant, no need for
rapid response
ncRNA: 1000’s of RNA’s, current
research
• miRNA’s - micro RNA hat
can degrade mRNA or block
translation
• Causes mRNA to fold on
itself and base pair to create
dsRNA which is then
digested with an enzyme
• Short interferring RNA
(siRNA) – also degrade
mRNA or block translation
(blocking by siRNA is called
RNAi, or RNA interferance)
 Protein Degradation
18_12ProteinDegradation_A.swf

• Proteosomes – break apart proteins in to

smaller peptide units

Chromatin changes

Transcription

RNA processing

Ubiquitin Proteasome
Translation
mRNA
degradation and ubiquitin
to be recycled
Protein processing Proteasome
and degradation

Protein to Ubiquinated Protein

be degraded protein fragments
(peptides)
Protein entering a
proteasome
 Single Gene Expression
• Different cells express different genes,
therefore they make different mRNA’s
• We can detect mRNA in a cell using nucleic acid
hybridization, by pairing it to a nucleic acid
probe
• Each probe is labeled with a fluorescent tag to
allow visualization
• The technique allows us to see the mRNA in
place (in situ) in the intact organism and is thus
called in situ hybridization
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
Figure 15.15-5

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
5 cDNA carries complete
5 3
coding sequence 3 5
without introns. cDNA
 Groups of Gene Expression
• Recall Microarray assays:
• Used to pinpoint differences in gene
expression between 2 different cell types
• How it’s done:
– Sequence a genome
– Use PCR to copy the genes (verification steps
here)
– Split the genes into single strands
– Place the single stranded DNA onto microscope
slides in spots (robots & computers do all this)