EDROLO CHAPTER 2E – GENE

REGULATION

AOS 1.1 The relationship between nucleic acids and Proteins

FROM VCAA:
• the basic elements of gene regulation: prokaryotic trp operon as a simplified
example of a regulatory process
 GENE REGULATION

• Gene regulation involves either inhibiting or activating gene expression.

• This prevents unnecessary production of gene products, saving energy.
• There are two types of genes involved in gene regulation:
• Structural genes:
• Produce proteins that are involved in the structure or function of the cell.
• Eg. Enzymes, transport proteins, peptide hormones
• Structural genes are found downstream of regulatory gene that controls them (toward the 3’ end)
• Regulatory genes:
• Produce regulatory proteins such as repressor proteins, which inhibit or decrease the expression of the
structural gene.
• Activator proteins initiate or increase the expression of structural genes.
• Can control the types of post-transcriptional modifications that occur to pre-mRNA (eg. Alternative splicing)
 OPERON
• In some organisms, such as prokaryotes, multiple structural genes that share a common purpose can be
arranged into groups so that their expression is efficiently controlled by a single promotor and operator.
• This is known as an operon.
• Because operators are always located downstream of the gene’s promoter region, they can be used as a
binding site for repressor or activator proteins.
Example 1:
• Operator region is bound with a repressor protein – RNA polymerase cannot move downstream from the promoter
region. Inhibiting transcription of the gene.
Example 2:
• Operator region is not bound with a repressor protein – RNA polymerase is free to move downstream from the
promoter region, allowing transcription of the gene to occur.

Example 1: Example 2:

Repressor
RNAPRepressor RNAP
protein
protein

Edrolo 2021
 THE trp OPERON

• The trp operon is present in some bacterial

species, such as E. coli.
• Regulates the expression of structural genes
which code for proteins involved in amino acid
tryptophan production.
• The trp operon consists of:
• Structural genes – trpE, trpD, trpC, trpB and
trpA
• Structural promoter – Site for RNA polymerase
to bind to transcribe structural genes
• Operator – Site for Repressor protein to bind
• Leader Region – Important for attenuation
• There is a regulatory gene located upstream of the
operon.
 TRYPTOPHAN

• The amino acid tryptophan can be used as a building block for complex proteins.
• To conserve energy, the structural genes are only expressed when tryptophan is
required.
• The expression of the trp structural genes depends on the level of tryptophan
present within the cell.
 GENE REGULATION BY REPRESSION

• Repression occurs when intracellular tryptophan levels are high.

High levels of Intracellular tryptophan: Low levels of Intracellular tryptophan:

Repression No Repression
 G E N E R E G U L AT I O N B Y R E P R E S S I O N :
H I G H TRYPTO P HAN L EVE LS

• Regulatory gene is constantly producing repressor protein

• Two tryptophan binds to repressor protein, inducing a conformational change which activates repressor
protein.
• Active repressor protein binds to operator region, blocking path of RNA polymerase.
• Transcription of the trp structural genes is repressed.
 G E N E R E G U L AT I O N B Y R E P R E S S I O N :
H I G H TRYPTO P HAN L EVE LS

RNA polymerase is blocked

by the repressor protein,
preventing transcription.

No transcription of structural
genes means no proteins
Two tryptophan bind to produced.
repressor protein, activating
it.
Repressor protein binds to
operator.

Edrolo 2021
 GENE REGULATION BY REPRESSION:
LOW TRYPTOPHAN LEVELS

No tryptophan bound to
repressor protein. Repressor
detaches from operator.
RNA polymerase is
Transcription free to transcribe
results in mRNA structural genes.
molecule which
will then go onto
translation to form
polypeptide chain
required to build
more tryptophan.
 ATTENUATION

• Prevents the completion of transcription.

• Occurs when there is no free tryptophan in the cell but still tryptophan present on tRNA molecules.

• No need to use energy and make more trp

• However, no trp bound to repressor protein so RNA polymerase can begin transcription.

• As RNA polymerase is transcribing, a ribosome binds to and translates at the mRNA at the same time.

• Both RNA polymerase and the ribosome move along at the same pace.
 GENE REGULATION BY ATTENUATION

Leader Region:
• Attenuator at the downstream end of leader region.
• A sequence of A – T bonds, transcribed into A – U bonds which are weak (only 2 hydrogen bonds)
• Four Regions
• Each region is partly complementary to the next (e.g. region 1 is partly complementary to region 2.
Region 2 is partly complementary to region 3 etc).
• Between region 1 and 2 there is a stop codon (on the transcribed mRNA).
• Region 1 contains 2 trp codons.
 GE N E R E GUL AT I ON B Y AT T E N UAT I ON.

• Attenuation occurs when the tRNA-bound tryptophan levels are high.

1. The processes of transcription and translation of

the trp operon begin and occur simultaneously.

2. The ribosome involved in translation arrives at

the attenuator sequence that codes for two
tryptophan amino acids. The tRNA-bound
tryptophan that is present in the cell travels to
the ribosome and is added to the protein that is
being made by the ribosome.
 GENE REGULATION BY ATTENUATION
CONTINUED…

3. This causes the mRNA molecule being read by the

ribosome to fold in a specific way via hydrogen bonds and
form a terminator hairpin loop.
4. The folding of the terminator hairpin loop causes the
mRNA molecule to separate from the template DNA at the
attenuator sequence.
5. RNA polymerase detaches from the DNA, causing
transcription to stop before any structural genes are
transcribed. Without these structural genes, new tryptophan
cannot be synthesised.
 Weak bonds in
attenuator break

Transcription
termination
signal

• The ribosome will continue translation until it reaches the stop codon, where it stops.
• RNA polymerase continues transcription until it reaches attenuator.
• Region 3 and 4 bind to form a hairpin loop (as they are partly complementary). This acts as the
transcription termination signal.
• As these regions bind to each other, they pull the mRNA strand from the DNA due to the weak bonds in
the attenuator region.
• This flings off the RNA polymerase from the DNA, leaving the structural genes untranscribed.
• This results in a short (attenuated) mRNA strand.
 G E N E R E G U L AT I O N B Y AT T E N U AT I O N C O N T I N U E D …

Low Levels of tRNA-bound tryptophan:

Note: not on the study design! You do NOT need to know this but helps to explain aspects of attenuation.
1. The processes of transcription and translation of the trp operon begin and occur simultaneously.
2. The ribosome involved in translation arrives at the attenuator sequence that codes for two tryptophan amino acids.
Due to there being no tRNA-bound tryptophan in the cell, when the ribosome involved in translation arrives at the
attenuator sequence that codes for two tryptophan amino acids it pauses. Meanwhile, the RNA polymerase
involved in transcription continues along the DNA.
3. This causes the mRNA molecule to fold in a specific way via hydrogen bonds and form an anti-terminator hairpin
loop.
4. The anti-terminator hairpin loop does not cause the mRNA to separate from the template strand at the attenuator
sequence.
5. RNA polymerase continues to read the DNA template strand, transcribing the structural genes for proteins
involved in the synthesis of tryptophan and translation can continue.
 N O AT T E N U AT I O N Note: not on the study design! You do NOT need to know this
but helps to explain aspects of attenuation.
• This occurs when there is no tryptophan in the cell at all.
• This time, the ribosome stops at the two trp codons. As there is no tryptophan on the tRNA molecules, translation cannot proceed.
• RNA polymerase continues transcription.
• As the ribosome has stopped at the trp codons and not the stop codon, it is no longer preventing regions 2 and 3 from binding and
forming a hairpin loop. This hairpin loop does not stop transcription.
• The bonds between DNA and RNA are stronger at region 4 than the attenuator, so the mRNA does not detach from DNA, and RNA
polymerase is left to continue transcribing the structural genes needed to produce more tryptophan.
• This results in a longer strand of mRNA.
 IMPORTANT INFORMATION ABOUT ATTENUATION
• You must be able to describe how attenuation occurs when tryptophan levels are high.
• You do not need to describe attenuation when tryptophan levels are low.
• You must understand the difference between repression (refers to intracellular trp) and attenuation
(refers to tRNA-bound trp).
• The processes of repression and attenuation are not perfect. A low level of transcription and
translation of the trp operon will occur regardless of the regulation. This is important as it ensures
that a cell is never completely without tryptophan. Still, repression and attenuation are critical for the
cell to function efficiently.
trp OPERON SUMMARY
 TASKS

Operon Activity Ch 2E MCQ Teams