Regulation of Gene Expression in

Prokaryotic and Eukaryotic Cells

Tri Panjiasih Susmiarsih

 Prokaryotic Regulation
• an operon is a functioning unit of DNA containing a cluster of
genes under the control of a single promoter
• Operon Segmen:
- Regulator Gene (upstream of rest of operon; recipe for a
protein called repressor protein)
- Promoter (where RNA Polymerase attaches)
- Operator Gene (“on/off” switch for operon)
- Structural Genes (contains recipes for proteins)
The parts of an operon
 Parts of an Operon

Regulator Gene
Promoter Structural Genes Terminator

DNA
When Repressor is at the operator,
Operator
RNA Polymerase can’t attach to the
RNA Repressor Promoter and transcription is blocked
Polymerase
There are 2 types of negative control operons:
 Inducible Operons
- are normally turned “off”
- an inducer turns them “on”
 Repressible Operons
- are normally turned “on”
- a repressor/corepressor turn “off”
 trp operon
• Bacteria need amino acids to build proteins, ex. tryptophan.
• E. coli can make their own tryptophan using enzymes that are
encoded by five genes, is called the trp operon.
• If tryptophan is present in the environment, E. coli bacteria
don't need to synthesize it, the trp operon is switched "off."
• When tryptophan availability is low, the operon is switched
"on," the genes are transcribed, biosynthetic enzymes are
made, and more tryptophan is produced.
• tryptophan binds to repressor, causing a change in shape,
tryptophan is a corepressor, repressor binds to operator
• therefore trp operon is a repressible operon
• The trp operon is a repressible operon (normally “on” but can
be turned “off”)
 The trp operon

Tryptophan is present, tryptophan bind the repressor protein at the operator sequence ----
blocks the RNA polymerase from transcribing the tryptophan genes.
tryptophan is absent, the repressor protein does not bind to the operator genes are
transcribed.
trp operon
This operon responds to the cell’s need for
the amino acid tryptophan.
This operon is usually turned “on” as the
cell needs a lot of trptophan.
trp operon

Tryptophan binds allosterically to repressor protein

 lac Operon
– The lac Operon
• usually in the “off” position
• three structural genes code for three enzymes involved in
lactose metabolism
– one breaks lactose into glucose and galactose
– another allows lactose to enter the cell
• the regulator codes for a repressor that ordinarily binds to
operon
• There are three genes in the lac operon that code for enzymes that aid
in the metabolism of lactose:
1. β-galactosidase (lacZ): converts lactose into glucose and galactose
2. β-galactoside permease (lacY): transports lactose into the cell
3. β-galactoside transacetylase (lacA): adds acetyl-group to galactose
(function still unknown)
 Prokaryotic Regulation
– The lac Operon
– regulation
– when lactose is absent, cell does not need
lactose-metabolizing enzymes
– when glucose is absent and lactose is present,
lactose binds to the repressor, causing a change
in shape
» lactose is an inducer
– repressor cannot bind to operator
– therefore lac operon is an inducible operon
– when lactose is present, the operon is “on” and
when it is absent, the operon is turned “off”.
The lac operon
When the inducer – lactose – is absent,
repressor protein binds to the operator
gene preventing RNA Polymerase from
transcribing the structural genes. The
operon is turned “off”.
 Prokaryotic Regulation
– The lac Operon, cont.
• cAMP/CAP system
– when glucose is present, more ATP is available (and less
cAMP)
– when glucose is absent, cyclic AMP (cAMP) accumulates
» cAMP: derived from ATP; has one phosphate
attached in two placescAMP binds to catabolite
activator protein (CAP)
– complex attaches to CAP binding site next to lac promoter
– DNA bends to better expose promoter to RNA polymerase,
increasing the rate of transcription
Cyclic AMP
CAP activity, no glucose
CAP activity, glucose
 Operon summary
Repressible operon
usually functions in anabolic pathways
synthesizing end products
when end product is present cell allocates
resources to other uses
Inducible operon
usually functions in catabolic pathways,
digesting nutrients to simpler molecules
produce enzymes only when nutrient is
available
cell avoids making proteins that have nothing to do
 Lac Operon - cAMP
– The lac Operon, cont.
• cAMP/CAP system, cont.
– when [lactose] = 0, no transcription
– when [lactose] = high and ATP:cAMP = high, minimum
transcription
– when [lactose] = high and ATP:cAMP = low, maximum
transcription
 Eukaryotic Regulation
• Eukaryotic DNA differs from prokaryotic DNA
Prokaryotes Eukaryotes
circular chromosomes linear chromosomes
one chromosome many chromosomes
no proteins many proteins
not repetitive highly repetitive
regulated by operons regulation more
complex
Control of Eukaryotic Gene
Expression
The control points of gene expression can occur at any step in
the pathway from gene to
functional protein
Eukaryotic gene regulation
 Eukaryotic Regulation
• Eukaryotes possess a variety of mechanisms to regulate
gene expression
– Chromatin Structure
1. chromatin consists of DNA (2 nm) wound around histones
– histones help organize DNA and prevent access to DNA
2. each core of eight histones (and DNA) forms a nucleosome (11 nm)
3. nucleosomes coil (30 nm)
4. euchromatin: looped chromatin (300 nm)
– state most chromatin is in
5. heterochromatin: condensed chromatin (700 nm)
– inactive
6. condensed chromosome (1,400 nm)
Levels of chromatin struct.
Hetero- and euchromatin
A nucleosome
 Eukaryotic Regulation
– Transcriptional Control
• no operons
• Transcription Factors and Activators
– transcription factors: proteins that regulate transcription
» bind to promoter
» attract and bind RNA polymerase
– transcription activators: also promote transcription
» bind to enhancer
– bridged by mediators

• Transposons
– transposons are DNA sequences that can move within
and between chromosomes
– they usually decrease or shut down gene expression
Initiation of transcription
Indian corn
 Eukaryotic Regulation
– Posttranscriptional Control
• mRNA processing
– introns removed and exons spliced
• speed with which mRNA leaves the nucleus
– Translational Control
• often involves 5’ cap or 3’ poly-A tail
– Posttranslational Control
• inc. protein activation or destruction
1.DNA packing
The chromosome (DNA molecule) is
wound around a group of 8
positively charged histone
proteins (bind tightly to negatively
charged DNA), is called a
nucleosome the basic unit of DNA
packing.
These nucleosomes resemble
beads on a string, further coiled
to produce a thicker structure
called chromatin
 Chromatin structure affects the
availability of genes for transcription

Some of the chromatin is tightly packed – it contains genes that

are seldom used (genes are not transcribed). This tightly
packed chromatin is called heterochromatin.
Genes that are transcribed (used) are more loosely packed into
areas that “loop” out – called looped domains – and form
what is known as euchromatin (true chromatin).
The final product of DNA packing is the chromosome
(shown here as a metaphase chromosome).
 2. Modifying the Chromatin
DNA Methylation – attaching methyl (CH3) groups turns
“off” genes. Example is Barr Body (extra X chromosome)
2. Modifying the Chromatin

Histone acetylation activates genes = on

• attachment of acetyl groups (–COCH3) to certain amino

acids of histone proteins neutralizes their positive charges
and they no longer bind to neighboring nucleosomes
• they change shape & grip DNA less tightly = unwinding DNA
• transcription proteins have easier access to genes
 3. Transcriptional Control

DNA must be unpacked

and if gene is methylated,
the methyl group must be
removed. Proteins called
transcription factors assist
in this job.
4. mRNA Processing

Introns must be cut out,

exons sewed together,
and a GTP cap and poly
A tail added.
Of course, the newly
formed mRNA can also
be destroyed at this
point if the cell has
changed its mind.
 5. mRNA Leaves Nucleus
A fully processed mRNA must
leave the nucleus in order to
be translated into protein.
The large nuclear pores must
be opened for its passage.
Again, the cell can change its
mind and not open the pores.
 Translation requires that ribosomes
6. Translation participate.
7.Post transcription Controls

Finally, the new protein

must be “folded” to
become active. The
cell can withhold this
folding or even
degrade (breakdown)
the protein at this
step.
Phosphorylation addition of phosphorous group on amino acods. either activates a protein or
deactivates it.
Glycosylation addition of monosaccharide on the protein residues. plays majhor role in various
membrane proteins. in some cases polysaccharides are also added.
Ubiquitination addition of ubiquitin on the protein which is majorly to tag protein for
degradation.
S-Nitrosylation added as S-nitrothiols usually on cysteines in the protein but is a random
process.
Methylation addition of methyl group on Nitrogen or oxygen of amino acid in the protein
N-Acetylation addition of acetyl group on nitrogen or oxygen of amino acid in the protein
Lipidation addition of lipids on proteins usually the membrane proteins are loaded with lipids.