Control of Prokaryotic and

Eokaryotik Gene Expression

MASYHUDI, DRG, MSI

BLOK 3 MODUL 4
PRODI GIGI UNMUL
 Prokaryotic Regulation of Genes
Regulating Biochemical
Pathway for Tryptophan
Synthesis.
1. Produce something that
will interfere with the
function of the enzyme
in the pathway.
2. Produce a gene
regulator that can inhibit
the transcription of one
biochemical pathway
enzymes.

2
1. Eukaryotic cells have many more
genes (i.e. 23,000 in human cells)
in their genomes than prokaryotic
cells (i.e. average 3000).
2. Physically there are more
obstacles to regulate eukaryotic
genes because there is so much
more DNA to manage. For
example, eukaryotic chromatin is
wrapped around histone proteins.
3. In addition there are other
nonhistone proteins that are used
in eukaryotic gene expression that
are not used in prokaryotic gene
3
 Operon and Prokaryotic Gene Expression

• Operon- Sekelompok gen prokariotik dengan

fungsi terkait yang sering dikelompokkan dan
ditranskripsikan bersama. Operon hanya
memiliki satu wilayah promotor untuk seluruh
operon.
4
 Operon and Prokaryotic Gene Expression

An operon is composed of the following:

• Structural genes- genes that are related and used in a
biochemical pathway.
• Promoter-The nucleotide sequence that can bind with
RNA polymerase to start transcription. This sequence
also contains the operator region.
• Operator-The nucleotide sequence that can bind with
repressor protein to inhibit transcription.
5
 Regulator Genes and Repressors
• Regulator gene- This gene produces a
protein called a repressor that can inhibit
the transcription of an operon by attaching
to the operator.

6
 Interaction of Modulators and Repressors

• Repressors have allosteric properties.

Modulators can bind to the repressor at an
allosteric site changing the conformation of the
repressor, thereby activating or deactivating the
repressor. Usually the modulator is a product of
the biochemical pathway.

7
Regulatory Proteins

8
Lactose and the Inducible lac Operon (negative
regulation)

Negative Gene Regulation

1. Inducible operon- the lac operon. This operon has
the ability to convert lactose into glucose and galactose.
This involves three structural genes

9
Animation of the lac Operon and Presence
of Lactose

10
Absence of Lactose and the lac Operon

• If no lactose or allolactose, mirip laktosa (Inducer) is

present, the repressor protein is active, binding to the
operator site. This prohibits the RNA polymerase from
transcribing the operon.
11
Animation of the lac Operon and Absence of
Lactose

12
 Synthesis of Tryptophan and the
Repressible trp Operon

Gen untuk enzim yang dibutuhkan untuk mensintesis triptofan

umumnya berkelompok dalam operon trp dan diekspresikan setiap
kali persediaan yang ada terbatas. 13
Dengan tidak adanya triptofan, protein repressor trp yang dikodekan
oleh operon terpisah, trpR, disintesis dan membentuk dimer.
Namun, ini tidak aktif sehingga tidak dapat mengikat ke operator trp
dan gen struktural dari operon trp ditranskripsi.

14
Animation of the trp Operon and Absence of
Tryptophan

15
Tryptophan Present and the Repressible trp Operon

Ketika triptofan ada, itu mengikat repressor trp dan menginduksi

perubahan konformasi pada protein itu, memungkinkannya untuk
mengikat operator trp dan mencegah transkripsi (operon
ditekan).
16
Pengikatan protein repressor ke operator tumpang tindih
dengan promotor, sehingga RNA polimerase tidak dapat
mengikat prometer. Oleh karena itu transkripsi dihentikan.
Dalam peran ini, triptofan dikatakan sebagai co-repressor.
Ini adalah kontrol negatif, karena repressor terikat
mencegah transkripsi.

17
Ketika triptofan hadir, enzim untuk biosintesis triptofan tidak diperlukan sehingga
ekspresi gen ini dimatikan.
Dicapai dengan mengikat triptofan ke repressor untuk mengaktifkannya sehingga
sekarang mengikat ke operator dan menghentikan transkripsi gen struktural.
Pengikatan protein repressor ke operator tumpang tindih dengan promotor, sehingga
RNA polimerase tidak dapat mengikat prometer. Sehingga transkripsi dihentikan.
Dalam peran ini, triptofan dikatakan sebagai co-repressor. Ini adalah kontrol negatif,
karena repressor terikat mencegah transkripsi. 18
Animation of the trp Operon and Presence of
Tryptophan

19
Lac and trp Operons-Examples
of Negative Gene Regulation
• The lac and typ operons are example of
negative gene regulation as the repressor
protein inhibits transcription of the
operons.
• Dalam regulasi negatif, protein repressor
mengikat operator untuk mencegah gen
diekspresikan.
20
Example of Positive Gene Regulation

Memerlukan
faktor
transkripsi
untuk mengikat
promotor agar
RNA
polimerase
memulai
transkripsi.

21
Both Lactose and Glucose Present

22
23
Gene expression of eukaryotic cells
• each cell maintain specific program /
differential gene expression
• one mRNA carries information for one gene
(monogennic mRNA)
• posttranscription modification of RNA
(removing introns and connecting exons)
• complicated regulation system, performed at
the several levels (transcription, translation,
protein activation + secretion)
 Complicated regulation system

• chromatin changes
• transcription
• processing RNA
• transport to cytoplasm
• degradation of mRNA
• translation
• cleavage, chemical modification
• protein degradation
Stages in gene expression in eukaryotic cell
1. Chromatin changes

• Heterochromatin is highly condensed -

transcriptional enzymes can not reach the DNA
• Acetylation / deacetylation of histons
• Methylation [cytosin] - inactive DNA is highly
methylated

DNA methylation and histone deacetylation repress

• DNA methylation
is esential for long-term inactivation of genes during
cell differentiation
Gene imprinting in mamals
• methylation constantly turns off the maternal or the
paternal allele of a gene in early development
• certain genes are expressed in a parent-of-origin-
specific manner
Epigenetic inheritance
 2. Transcription

• proteins that bind to DNA and facilitate of inhibit

binding of RNA polymerase

• transcription initiation complex
• transcription factors – general transcription
factors
for all protein-coding genes
- specific transcription factors – transcription of
particular genes at the appropriate time and place
-
Eukaryotic gene and transcript
• Cell-type specific transcription

• Genes coding for the enzymes of a metabolic

pathway are scattered over different
chromosomes - coordinated control in
response of chemical signals from outside
the cell - receptors
signal transduction pathways activating of
transcription activators or repressors
Signal transduction pathways
3. Processing RNA

Post-transcriptional modifications
Alternative splicing
The same primary transcript, but different the
mRNA molecule / exons and introns

4, 5. transport of mRNA / degradation

Lifespan of mRNA is important for protein synthesis

Enzymatic shortening
6. Translation
At the initiation stage –
regulatory proteins that bind at the 5’ end of the
mRNA
Activation or inactivation of protein factors to initiate
translation
7. Cleavage, chemical modification

Cleavage

Post-translational modifications
Regulatory proteins [products] are activated
or inactivated by the reversible addition of
phosphate groups / phosphorylation
Sugars on surface of the cell / Glycosylation
• Polypeptide chain may
be cleaved into two or
three pieces
• Preproinsulin
• Proinsulin - disulfide
bridges
• Insulin
• Secretory protein
Post-translational modifications

Acid/base - act/inact
Hydrolysis – localization, act/inact
Acetylation - act/inact
Phosphorylation - act/inact
Prenylation - localization
Glycosylation - targeting
Various steps in the synthesis and
assembly of collagen fibrils
8. protein degradation
Lifespan of protein is strictly regulated
Marked protein for destruction is attached by a small protein ubiquitin
Protein complexes proteasomes
Proteasomes : protein complexes which degrade unneeded or damaged proteins by
proteolysis, a chemical reaction that breaks peptide bonds. Enzymes that help such
reactions are called proteases.

Proteasomes are found inside all eukaryotes

and archaea, and in some bacteria. In
eukaryotes, proteasomes are located both in the
nucleus and in the cytoplasm.