GENE EXPRESSION REGULATION

of PROCARYOTES & VIRUSES

Fera Ibrahim & T. Mirawati Sudiro

DEPARTEMEN MIKROBIOLOGI FKUI

Details of The process are different in
eucaryotes/procaryotes !
 GENE EXPRESSION
CONSTITUTIVE GENE EXPRESSION !
genes that are always active
genes that are always "turned on"
genes are always needed
eg: genes that code for enzymes of glycolytic pathway
Constitutive genes
cellular housekeeping functions
tRNA, rRNA, ribosomal protein, RNA polymerase subunit

INDUCIBLE OR REPRESSIBLE GENE EXPRESSION

INDUCTION
enzymes for catabolic pathways (degradation)
synthesized only when needed (Expression occurs only
when substrate enzymes present ) induction
REPRESSION
enzymes from anabolic pathways (synthesis)
If product presents, gene expression turn off repression
( ADAPTIVE)
INDUCIBLE AND REPRESSIBLE GENE EXPRESSION
!
REGULATION OF GENE
EXPRESSION IN
PROKARYOTES
GENE ORGANIZATION
OF PROKARYOTES
REGULATORY GENE

OPERON :
!
The gene cluster and promoter,
plus additional sequences
that function together in regulation
Promotor
Operator
Structural Gene

Jacob and Monod proposed the Operon Model for gene regulation
 Gene Regulation in Prokaryotes
Bacteria have a simple general mechanism for coordinating
the regulation of genes that encode products involved in a set
!
of related processes.

A. some genes not regulated

-constitutive genes = genes that are always active
-genes that are always "turned on"
-genes are always needed
-eg: genes that code for enzymes of glycolytic pathway
B. some genes are regulated
-some genes only needed under certain conditions
-transcription can be "turned on or off"
-enzymes produced from these genes vary greatly in
cytoplasmic concentration
 Types of regulation
A. Inducible operon
!
- Induction is associated with catabolic pathways
-enzymes for a given catabolic pathway synthesized only when needed
-eg: catabolism of lactose

B. Represible operon
- Repression is associated with anabolic pathways
-focus on the "end products" of anabolic pathways
-amount enzyme varies inversely with amount of end product in cell
- This operon is normally in on mode, and will be turned off only when the end
product is no longer required
- Excess product plays a role as a corepressor, that slows the transcription of the
operon
-eg: synthesis of amino acids, purines and pyrimidines in cell
 !
A. In the absence of lactose:
- A repressor ataches to the operator of
the operon --- locks the operator ---
suppress transcription of structural
proteins downstream of it
(OPERON OFF)

B. In the presence of lactose

- Lactose as genetic inducer ---
attaches to the receptor inactive
repressor released from the operator
- RNA polymerase bind to the
promoter and initiate transcription
Repressible operon !
Example: regulation of arginine synthesis

A. Operon On
- A repressible operon remains on when its
nutrient pruduct (here: arginine) are in
great demand by the cells

B. Operon Off.
The operon is repressed when:
- Arginine builds up --- as a corepressor -
-- activates the receptor
- The repressor complex binds to the
operator ---- block RNA polymerase ---
transcription blocked
 ATTENUATION
!
 !
Attenuation

- Attenuation regulates the termination of transcription as a !

function of tryptophan concentration.
- At low levels of trp full length mRNA is made, at high levels
transcription of the trp operon is prematurely halted. !
- Attenuation works by coupling transcription to translation. !
- Prokaryotic mRNA does not require processing and since
prokaryotes have no nucleus, translation of mRNA can start
before transcription is complete. !
- Consequently regulation of gene expression via attenuation is
unique to prokaryotes. !
 !
a. Attenuation is mediated by the formation of one of two
possible stem-loop structures in a 5' segment of the trp operon in
the mRNA.

b. If tryptophan concentrations are low then translation of the leader !

peptide is slow and transcription of the trp operon outpaces translation.
This results in the formation of a nonterminating stem-loop structure
between regions 2 and 3 in the 5' segment of the mRNA. Transcription
of the trp operon is then completed.

c. If tryptophan concentrations are high the ribosome quickly translates

the mRNA leader peptide. !
Because translation is occurring rapidly the ribosome covers region 2
so that it can not attach to region 3.
Consequently the formation of a stem-loop structure between regions
3 and 4 occurs and transcription is terminated.
 Transformation is the alteration of a
! bacterial cells genotype by the uptake of
naked, foreign DNA from the surrounding
environment.
For example, harmless Streptococcus
pneumoniae bacteria can be transformed to
pneumonia-causing cells.
This occurs when a live nonpathogenic cell takes
up a piece of DNA that happens to include the
allele for pathogenicity from dead, broken-open
pathogenic cells.
The foreign allele replaces the native allele in the
bacterial chromosome by genetic recombination.
The resulting cell is now recombinant with DNA
derived from two different cells.
 Transduction occurs when a phage
! carries bacterial genes from one host cell
to another.
In generalized transduction, a small
piece of the host cells degraded DNA is
packaged within a capsid, rather than the
phage genome.
When this phage attaches to another
bacterium, it will inject this foreign DNA into its
new host.
Some of this DNA can subsequently replace the
homologous region of the second cell.
This type of transduction transfers bacterial
genes at random.
 CONJUGATION !
Conjugation transfers genetic material between
two bacterial cells that are temporarily joined.
One cell (male) donates DNA and its mate
(female) receives the genes.
A sex pilus from the male initially joins the two
cells and creates a cytoplasmic bridge between
cells.
Maleness, the ability to form
a sex pilus and donate DNA,
results from an F factor as a
section of the bacterial
chromosome or as a plasmid.
! Antibiotic resistance transfer
In the 1950s, Japanese physicians began to notice
that some bacterial strains had evolved antibiotic
resistance.
The genes conferring resistance are carried by plasmids,
specifically the R plasmid (R for resistance).
Some of these genes code for enzymes that specifically
destroy certain antibiotics, like tetracycline or ampicillin.
When a bacterial population is exposed to an
antibiotic, individuals with the R plasmid will
survive and increase in the overall population.
Because R plasmids also have genes that encode
for sex pili, they can be transferred from one cell
to another by conjugation.
! TRANSPOSON

A transposon is a piece of DNA that can move

from one location to another in a cells genome.
Transposon movement occurs as a type of
recombination between the transposon and
another DNA site, a target site.
In bacteria, the target site may be within the
chromosome, from a plasmid to chromosome (or vice
versa), or between plasmids.
Transposons can bring multiple copies for
antibiotic resistance into a single R plasmid by
moving genes to that location from different
plasmids.
This explains why some R plasmids convey resistance to
many antibiotics.
 Composite transposons (complex
transposons) include extra genes
sandwiched between two insertion
sequences.
It is as though two insertion sequences
happened to land relatively close together and
now travel together, along with all the DNA
! between them, as a single transposon.