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BIOLOGY 1A03
Cellular and Molecular Biology

Theme 3
Module 3 & 4 Review
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Module 3

Eukaryotic transcriptional regulation

 Promoters recruit transcription factors and 3

RNA polymerase

Unlike prokaryotes,
eukaryotic genes
ARE NOT clustered
into operons!

Instead, each gene

is regulated by its
own promoter and
regulatory regions
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Trans-acting factors interact specifically with DNA
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Eukaryotic transcription is regulated

Interruption of any of
these interactions can be
a means of
transcriptional
Regulatory regions regulation of gene
expression.
are often quite far
away from the core
promoter regions
in eukaryotes
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Transcriptional repressors can halt transcription
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Transcriptional repressors can halt transcription

X
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Transcriptional regulators determine cell types

ALL CELLS have

the same DNA
blueprint!

BUT- In some
cells, certain
genes are
activated, in
others, certain
genes are kept off
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In situ studies in gene expression

i.e
Target mRNA Fluorescent complementary probe
5’ 3’ 3’ 5’
...A C U U A C C C G G G A C U A... ...T G A A T G G G C C C T G AT...
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Identifying differences in gene expression

A cancerous cell is not a foreign cell to anyone’s body. It is actually a cell from a person’s
body that is just “misbehaving by having inappropriate regulation of gene expression
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Identifying differences in gene expression
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Preparing fluorescently labelled cDNA
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Using a microarray chip to measure gene expression
Microarray chips are as small as a glass slide and contain multiple wells
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Using a microarray chip to measure gene expression
Each well represents a gene of interest
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Using a microarray chip to measure gene expression
Each well represents a gene of interest
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Using a microarray chip to measure gene expression
In a microarray chip, each well is “pre-seeded” with single-stranded DNA that is anchored
to the bottom of the well, which is unique to each gene of interest

Gene 1 Gene 2 Gene 3

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Labelled cDNA hybridizes with DNA on chip
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Fluorescence analysis of gene activity
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Fluorescence analysis of gene activity

This well This well

This well
fluoresces fluoresces
fluoresces
in red in yellow
in green
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Interpreting our results
Eg. kills cells when
Eg. helps cancer they malfunction Eg. a gene that Eg. a gene that helps
cells move “cell suicide” codes for ribosomes cells become bone
apoptosis

Gene 1 Gene 2 Gene 3 Gene 4 (Control)

On in cancer cells On in normal cells

Off in normal cells Off in cancer cells On in both cell types
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Levels of gene expression regulation

Similar to prokaryotes,
environmental factors
(either around or
between cells, or from the
environment itself), can
influence and can alter
eukaryotic gene
expression at all levels
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Regulating eukaryotic gene expression
1. Histone Modifications
• Regulating the amount of mRNA that is transcribed
Chemical DNA modifications that occur along the
genome that change gene activity, but to not alter the
2. DNA methylation DNA sequence
• Regulating the amount of mRNA that is transcribed
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Regulating eukaryotic gene expression
1. Histone Modifications
• Regulating the amount of mRNA that is transcribed
Chemical DNA modifications that occur along the
genome that change gene activity, but to not alter the
2. DNA methylation DNA sequence
• Regulating the amount of mRNA that is transcribed

3. mRNA stability
• Regulating the amount of mRNA that is translated into proteins
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Regulating eukaryotic gene expression
1. Histone Modifications
• Regulating the amount of mRNA that is transcribed
Chemical DNA modifications that occur along the
genome that change gene activity, but to not alter the
2. DNA methylation DNA sequence
• Regulating the amount of mRNA that is transcribed

3. mRNA stability
• Regulating the amount of mRNA that is translated into proteins

4. Protein stability
• Regulating the amount of time a protein can function in a cell
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Regulating eukaryotic gene expression
1. Histone Modifications
• Regulating the amount of mRNA that is transcribed
Chemical DNA modifications that occur along the
genome that change gene activity, but to not alter the
2. DNA methylation DNA sequence
• Regulating the amount of mRNA that is transcribed

3. mRNA stability
• Regulating the amount of mRNA that is translated into proteins

4. Protein stability
• Regulating the amount of time a protein can function in a cell
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DNA compaction regulates gene expression
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Histone modifications alter DNA winding
1. Positively charged tails of
nucleosomal histone
proteins probably interact
with negatively charged
phosphates of DNA.
Acetylation

2. Acetylation of the tails

weakens their interaction
with DNA and may permit
some transcription factors
to bind to DNA.
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Methylation and acetylation of histone proteins
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Regulating eukaryotic gene expression
1. Histone Modifications
• Regulating the amount of mRNA that is transcribed

2. DNA methylation
• Regulating the amount of mRNA that is transcribed

3. mRNA stability
• Regulating the amount of mRNA that is translated into proteins

4. Protein stability
• Regulating the amount of time a protein can function in a cell
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Methylation near the promoter of a gene
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Methylation near the promoter of a gene
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Methylation promotes chromatin remodeling

Heavily methylated promoters

also recruit histone deacetylase
which is able to deacetylate or
remove acetyl groups from the
histone proteins tails…inhibiting
gene expression at the
transcriptional level
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Module 4

Turning off gene expression

(mRNA and protein stability)

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Regulating eukaryotic gene expression
1. Histone Modifications
• Regulating the amount of mRNA that is transcribed

2. DNA methylation
• Regulating the amount of mRNA that is transcribed

3. mRNA stability
• Regulating the amount of mRNA that is translated into proteins

4. Protein stability
• Regulating the amount of time a protein can function in a cell
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miRNAs result in inhibition of translation
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siRNAs lead to mRNA degradation
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Regulating eukaryotic gene expression
1. Histone Modifications
• Regulating the amount of mRNA that is transcribed

2. DNA methylation
• Regulating the amount of mRNA that is transcribed

3. mRNA stability
• Regulating the amount of mRNA that is translated into proteins

4. Protein stability
• Regulating the amount of time a protein can function in a cell
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Protein degradation

Proteasomes degrade unneeded or damaged proteins.

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Protein degradation
Proteins are removed from the cell by marking them for enzymatic degradation
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Protein degradation
Proteins are removed from the cell by marking them for enzymatic degradation
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Tomorrow’s applied lecture…

Epigenetics and Gene Regulations

(These two mice are genetically identical, and from the same litter!)