Gene expression in eukaryotes is regulated through various mechanisms including histone modification, DNA methylation, enhancers, and combinations of regulatory proteins. Gene regulation occurs through positive and negative elements that increase or decrease expression. Key examples are the lac and tryptophan operons in prokaryotes, which are regulated by repressors and inducers to control metabolic pathways in response to environmental conditions.
Gene regulation in eukaryotes in a nutshell covering all the important stages of gene regulation in eukaryotes at transcriptional level, translation level and post-translational level.
Gene regulation in eukaryotes in a nutshell covering all the important stages of gene regulation in eukaryotes at transcriptional level, translation level and post-translational level.
REGULATION OF
GENE EXPRESSION
IN PROKARYOTES & EUKARYOTES .
This presentation is enriched with lots of information of gene expression with many pictures so that anyone can understand gene expression easily.
Gene expression is the process by which the information encoded in a gene is used to direct the assembly of a protein molecule.
Gene expression is explored through a study of protein structure and function, transcription and translation, differentiation and stem cells.
It is the process by which information from a gene is used in the synthesis of a functional gene product.
These products are often proteins, but in non-protein coding genes such as ribosomal RNA (rRNA), transfer RNA (tRNA) or small nuclear RNA (snRNA) genes, the product is a functional RNA.
The process of gene expression is used by all known life - eukaryotes (including multicellular organisms), prokaryotes (bacteria and archaea)
Regulation of gene expression:
Regulation of gene expression includes a wide range of mechanisms that are used by cells to increase or decrease the production of specific gene products (protein or RNA).
Gene regulation is essential for viruses, prokaryotes and eukaryotes as it increases the versatility and adaptability of an organism by allowing the cell to express protein when needed.
CLASSIFICATION OF GENE WITH RESPECT TO THEIR EXPRESSION:
Constitutive ( house keeping) genes:
Are expressed at a fixed rate, irrespective to the cell condition.
Their structure is simpler.
Controllable genes:
Are expressed only as needed. Their amount may increase or decrease with respect to their basal level in different condition.
Their structure is relatively complicated with some response elements.
TYPES OF REGULATION OF GENE:
positive & negative regulation.
Steps involving gene regulation of prokaryotes & eukaryotes.
Operon-structure,classification of mechanisms- lac operon,tryptophan operon ,
and many things related to gene expression.
This is a video slide so anyone can understand this topic easily by seeing pictures included in this slide.
REGULATION OF
GENE EXPRESSION
IN PROKARYOTES & EUKARYOTES .
This presentation is enriched with lots of information of gene expression with many pictures so that anyone can understand gene expression easily.
Gene expression is the process by which the information encoded in a gene is used to direct the assembly of a protein molecule.
Gene expression is explored through a study of protein structure and function, transcription and translation, differentiation and stem cells.
It is the process by which information from a gene is used in the synthesis of a functional gene product.
These products are often proteins, but in non-protein coding genes such as ribosomal RNA (rRNA), transfer RNA (tRNA) or small nuclear RNA (snRNA) genes, the product is a functional RNA.
The process of gene expression is used by all known life - eukaryotes (including multicellular organisms), prokaryotes (bacteria and archaea)
Regulation of gene expression:
Regulation of gene expression includes a wide range of mechanisms that are used by cells to increase or decrease the production of specific gene products (protein or RNA).
Gene regulation is essential for viruses, prokaryotes and eukaryotes as it increases the versatility and adaptability of an organism by allowing the cell to express protein when needed.
CLASSIFICATION OF GENE WITH RESPECT TO THEIR EXPRESSION:
Constitutive ( house keeping) genes:
Are expressed at a fixed rate, irrespective to the cell condition.
Their structure is simpler.
Controllable genes:
Are expressed only as needed. Their amount may increase or decrease with respect to their basal level in different condition.
Their structure is relatively complicated with some response elements.
TYPES OF REGULATION OF GENE:
positive & negative regulation.
Steps involving gene regulation of prokaryotes & eukaryotes.
Operon-structure,classification of mechanisms- lac operon,tryptophan operon ,
and many things related to gene expression.
This is a video slide so anyone can understand this topic easily by seeing pictures included in this slide.
Gene regulation is the process used to control the timing, location and amount in which genes are expressed. The process can be complicated and is carried out by a variety of mechanisms, including through regulatory proteins and chemical modification of DNA.
presented by HAFIZ M WASEEM
university of education LAHORE Pakistan
i am from mailsi vehari and studied in lahore
bsc in science college multan
msc from lahore
This presentation provide knowledge about Gene Expression & its regulation in brief.
i hope it gives some information about gene expression in your academic time.
In this presentation mentioned - Lac Operon and its expressor.
Most bacteria are free-living organisms that grow by increasing
in mass and then divide by binary fission.
Growth and division are controlled by genes, the expression
of which must be regulated appropriately. Genes
whose activity is controlled in response to the needs of a
cell or organism are called regulated genes. All organisms
also have a large number of genes whose products
are essential to the normal functioning of a growing and
dividing cell, no matter what the conditions are. These
genes are always active in growing cells and are known as
constitutive genes or housekeeping genes; examples include
genes that code for the enzymes needed for protein
synthesis and glucose metabolism. Note that all genes are
regulated on some level. If normal cell function is impaired
for some reason, the expression of all genes, including
constitutive genes, is reduced by regulatory
mechanisms. Thus, the distinction between regulated
and constitutive genes is somewhat arbitrary.
The following topics are discussed
. Prokaryotic gene expression and regulation
Prokaryotic “gene structure”
The basic structure of Operon
Lactose Operon” regulation
Tryptophan Operon” regulation
2. Eukaryotic gene expression and regulation
Eukaryotic gene structure
Regulons
Imagine a situation when a cell starts producing enzymes required for metabolism and those required for cell death (apoptosis) at the same time. The cell will be in a confused state and will not know which function to perform first. The needs of the body keep changing with time and cell has to tune itself to perform the desired set of activities. Gene regulation helps a unicellular organism to adapt well to the environment.
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5. Negative regulation
A decrease in the gene expression due to the
presence of a regulatory element ( negative
regulator)
6. Genes
The genes are generally considered under
two categories
1) Constitutive genes
2) Inducible genes
7. Constitutive gene
The products (proteins) of these genes are
required all the time in a cell
The constitutive genes called house keeping
genes
They are expressed at more or less constant
rate in almost all the cells
Eg: the enzymes of citric acid cycle
8. Inducible gene
The concentration of the proteins synthesized
by inducible genes is regulated by various
molecular signals.
An inducer increases the expression of these
genes
Repressor decreases the expression of these
genes.
9. One cistron-one subunit concept
The cistron is the smallest unit of genetic
expression
One gene-one enzyme hypothesis is replaced
by one cistron-one subunit.
10. The operon concept
Introduced by Jacob and Monad in 1961
Based on their observation on the regulation
of lactose metabolism
It is popularly known as lac operon
11.
12. Structure of Lac operon
It consists of a regulatory gene
Regulatory gene I -for inhibition
Operator gene-O
Three structural gene-Z, Y, A
Promoter site next to operator gene.
Structural genes Z, Y, A-codes for the enzymes β-
Galactosidase-hydrolyses lactose (β-Galactoside)
to galactose and glucose
Permease is reponsible for the transport of
lactose into the cell
An mRNA coding for more than one protein is
known as polycistronic mRNA.
13. Repression of lac operon
Regulatory gene I is constitutive
Lac repressor is tetrameric regulatory protein
specifically binds to the operator gene (O)
This prevents the binding of the enzyme RNA
polymerase to the promoter site (P)
It blocks the transcription of structural genes
(Z,Y,A)
It happens in the absence of lactose in E.Coli
The repressor molecule act as a negative
regulator of gene expression
14. Derepression of lac operon
In the presence of lactose (inducer)- in the medium –a
small amount can enter the E.coli.
The repressor molecule have high affinity for lactose.
The lactose molecule bind and induce a conformational
change in the repressor.
Repressor gets inactivated and cannot bind to the
operator gene (O)
The RNA polymerase attaches to the DNA at the
promoter site and transcription proceeds
Formation of polycistronic mRNA (genes Z, Y and A)
and finally enzymes-β galactosidase, galactoside
permease and galactoside acetylase.
Lactose acts by inactivating the repressor molecule.
15. Gratuitous inducers
Structural analogs of lactose which can
induce the lac operon.
They are not the substrates for the enzyme β
galactosidase.
Such substances are known as gratuitous
inducers
Eg: IPTG-isopropylthiogalactoside
16. The catabolite gene activator
protein
The cells of E.coli utilize glucose in
preference to lactose when both of them are
present in the medium
The utilization of lactose starts after the
depletion of glucose.
Glucose somehow interferes with the
induction of lac operon.
The attachment of RNA polymerase to the
promoter site requires the presence of a
catabolite gene activator protein (CAP) bound
to cAMP.
17.
18. Continued..
The presence of glucose lowers the
intracellular concentration of cAMP by
inactivating enzyme adenylyl cyclase
Formation of CAP-cAMP is low
Binding of RNA polymerase to DNA and the
transcription is negligible in the presence of
glucose.
Glucose interferes with the expression of lac
operon.
CAP-cAMP act as a positive regulator for
gene expression.
19. Tryptophan operon
Is an aromatic aminoacid required for the synthesis of
all proteins that contain tryptophan.
If tryptophan is not present in the medium in adequate
quantity, the bacterial cell has to make it as it is
required for the growth of the bacteria
The operon contains five structural genes (trpE, trpD,
trpC, trpB, trpA) and
The regulatory elements-primary promoter (trpP),
operator (trpO), attenuator (trpa), secondary internal
promoter (Trp P2) and
Terminator (trpt).
20.
21. Continued..
The five structural genes of tryptophan operon
code for three enzymes (two enzymes contain
two different subunits)
The tryptophan repressor is always turned on,
unless it is repressed by a specific molecule
called corepressor.
Lactose operon is inducible whereas
tryptophan operon is repressible.
The tryptophan operon is said to be
depressed when it is actively transcribed.
22. Tryptophan operon regulation by
a repressor
Tryptophan act as a corepressor to shut down
the synthesis of enzymes from tryptophan
operon.
Brought in association with a specific protein
namely tryptophan repressor.
Tryptophan repressor, a homodimer binds
with two molecules of tryptophan and then
binds to the trp operator to turn off the
transcription.
It also regulates the transcription of the gene
(trpR) responsible for its own synthesis.
23. Continued..
Two polycistronic mRNAs are produced from
tryptophan operon
One derived from all the five structural genes.
Other obtained from the last three genes.
Tryptophan can also inhibit the enzyme
anthranilate synthetase besides acting as a
corepressor
This is referred to as feedback inhibition
Brought by binding of tryptophan at an
allosteric site on anthranilate synthetase.
24. Attenuator-second control site for
trytophan operon.
Attenuator gene (trpa) of tryptophan operon lies
upstream of trpE gene.
It is the second level regulation of tryptophan
operon.
The attenuator regions provides RNA polymerase
which regulates transcription.
In the presence of tryptophan-transcription is
prematurely terminated at the end of attenuator
region.
In the absence of tryptophan, the attenuator
region has no effect on transcription
Therefore the polycistronic mRNA of the five
structural genes can be synthesized.
25. Gene expression in eukaryotes
The gene expression in eukaryotes is
regulated to provide the appropriate response
to biological needs
a) Expression of certain genes (house keeping
genes) in most of the cells
b) Activation of selected genes upon demand
c) Permanent inactivation of several genes in
all but few types.
26. Continued..
In prokaryotic cells, most of the DNA is
organized into genes which can be
transcribed.
In mammals very little of the total DNA is
organized into genes.
The function of the bulk of the extra DNA is
not known.
27. Histone acetylation and
deacetylation
Eukaryotic DNA segments are wrapped around histone
proteins to form nucleosome.
Acetylation or deacetylation of histones is an important
factor in determining the gene expression
Acetylation of histones leads to activation of gene
expression while deacetylation reverses the effect.
Acetylation predominantly occurs on the lysine
residues in the amino terminal ends of histones
The modification in histones reduces the positive
charges of terminal ends (tail), and decreases their
binding affinity to negatively charged DNA.
So the nucleosome structure is disrupted and and it
allow easy transcription.
28. Methylation of DNA and
inactivation of genes
Cytosine in the sequence CG of DNA gets
methylated to form 5’-methylcytosine
Methylation leads to loss of transcriptional activity
and thus inactivation of genes
This occurs due to binding of methylcytosine
binding proteins to methylated DNA.
The result is methylated DNA is not exposed and
bound to transcription factors
29.
30. Enhancers and tissue specific
gene expression
Enhancers (or activators are DNA elements
that facilitate or enhance gene expression
The enhancers provide binding sites for
specific proteins that regulate transcription
They facilitate binding of the transcription
complex to promoter regions.
31. Difference between enhancers
and promoters
Enhancers located thousands of base pairs away from
the start of transcription site-promoters close to the site
of transcription
They can work in either orientation i.e. enhancers can
work upstream (5’) or downstream (3’) from the
promoter.
Some enhancers possess the ability to promote
transcription in a tissue specific manner.
Gene expression in lymphoid cells for the production of
immunoglobins (Ig) is promoted by the enhancer
associated with Ig genes between J and C regions.
Tissue specific gene expression is largely mediated
through the involvement of enhancers.
32. Combination of DNA elements and
proteins in gene expression
Mammalian gene expression is a complicated process
with several environmental stimuli on a single gene.
Ultimate response-combination of DNA and proteins
Gene I is activated by combination of activators 1,2
and 3
Gene II is more effectively activated by the combined
action of 1,3 and 4
Activator 4 is not in direct contact with DNA, but it
forms a bridge between activators 1 and 3 and
activates gene II
33.
34. Continued..
Gene III gets inactivated by a combination of
1, 5 and 3
In this case, protein 5 interferes with the
binding of protein 2 with the DNA and
inactivates the gene.
35. Motifs in proteins and gene
expression
Literally means dominant element
Motifs in proteins mediate the binding of
regulatory proteins (transcription factors) to
DNA.
The specific control of transcription occurs by
the binding of regulatory proteins with high
affinity to the correct regions of DNA
36. Continued..
Protein DNA interactions are brought out by
four unique motifs.
a) Helix-turn-helix (HTH)
b) Zinc finger
c) Leucine zipper
d) Helix-loop-helix (HLH)
Motif DNA interactions are maintained by
hydrogen bonds and van der Waals forces.
37. Helix turn helix (HTS)
Motif is about 20 aminoacids which
represents a small part of large protein.
HTS is the domain part of the protein which
specifically interacts with the DNA.
Eg: lactose repressor, cyclic AMP catabolite
activator protein (CAP) of E.coli.
Homeodomain proteins- portion of the protein
of the transcription factors in mammals that
recognises DNA.
38.
39. Zinc finger motif
The transcription factors TFIIIA requires zinc
for its activity.
TFIIIA contains zinc ions as a repeating
coordinated complex.
The complex is formed by the closely spaced
aminoacids cysteine and cysteine, followed by
a histidine-histidine pair.
His His is replaced by a second Cys-Cys pair.
40. Continued..
The zinc fingers bind to the major groove of
DNA, and lie on the face of the DNA.
The binding makes a contact with 5 bp of
DNA.
The steroid hormone receptor transcription
factors use zinc motifs to bind to the DNA.
Mutation→single aminoacid change of zinc
finger→lead to resistance to the action of
certain hormones on gene expression.
Mutated zinc finger→resistant to the action of
calcitriol→result in rickets.
41.
42. Leucine zipper motif
The basic region of leucine zipper (bZIP)
proteins are rich in the aminoacid leucine.
Occurs as a periodic repeat of leucine
residues at every seventh position.
Repeat structure allows two identical
monomers or heterodimers to zip together
and form a dimeric complex.
This protein protein complex associates and
interacts with DNA.
Eg: leucine zipper proteins are enhancer
binding proteins (EBP)-fos and jun.
43.
44. Helix loop helix motif
Two amphipathic (feeling of closeness) α-
helical segments of proteins can form helix-
loop-helix motif and bind to DNA.
The dimeric form of protein bind to the DNA
45.
46. Gene regulation in eukaryotes
Eukaryotic cells employ variety of
mechanisms to regulate gene expression
1) Gene amplification
2) Gene rearrangement
3) Processing of RNA
4) Alternate mRNA splicing
5) Transport of mRNA from nucleus to
cytoplasm
6) Degradation of mRNA.
47. Gene amplification
The expression of a gene is increased several
fold.
The malignant cells develop drug resistance
to long term administration of methotrexate by
amplifying the genes coding for dihydrofolate
reductase.
48. Gene rearrangement
Responsible for molecular mechanism of antibody
diversity
Transposition of genes and somatic recombination of
DNA-basis of gene rearrangement
The structure of immunoglobin molecule contains two
light chain (L) and two heavy chains(H)
Each of these chains (Lor H) contains an N terminal
variable and C terminal constant (C) regions
V regions of immunoglobin are responsible for the
recognition of antigens
Each light chain can be synthesized by three distinct
DNA segments-VL, joining JL, and the constant CL
500 VL segments, 6 JL segments and 10 CL segments.
49.
50. Continued..
Formation of heavy chain occurs by the
rearrangement of 4 distinct genes-variable
(VH), diversity (D), joining (JH) and constant
(CH)
51. Processing of RNA
The changes include intron-exon splicing,
polyadenylation
52. Alternate mRNA splicing
Different mRNAs can be produced by
alternate splicing which code for different
proteins.
53. Degradation of mRNA
The expression of genes is indirectly influenced by
the stability of mRNAs.
The ends of mRNA determines the stability of
mRNA.
Eukaryotic mRNA has 5’ non coding sequences
(5’-NCS), a coding region and a 3’-NCS.
All mRNA are capped at the 5’ end polyadenylate
sequence at the 3’ end.
The 5’ cap and poly (A) tail protect the mRNA
against the attack by exonuclease
Stem loop structures in the NCS region and AU
rich regions in the 3’ NCS provide stability to
mRNA.