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2. CONTENTS
●PROMOTERS
●ROLE IN TRANSCRIPTION
●GENERAL TYPES OF PROMOTERS
● PROMOTER REGIONS
● PROKARYOTIC PROMOTERS
● EUKARYOTIC PROMOTERS
● BIDIRECTIONAL PROMOTERS
● SUBGENOMIC PROMOTERS
● DETECTION OF PROMOTERS
● MOST COMMONLY USED PROMOTERS
● THE AMAZING T7 PROMOTER
● OPERONS
● STRENGTH OF PROMOTER
● DISEASES ASSOCIATED WITH ABERRANT PROMOTER
● WHY INTEREST IN PROMOTERS
● REFERENCES
3. What causes the alleles in the genes
to be turned off in certain areas of
the body where they are not
needed?
-A high school student from California
March 20, 2008
Great question!
A big part of the answer can be
found in “Promoters."
4. A promoter is the main regulatory
portion of a gene. The simplest
analogy is that a promoter is a
“switch” that turns a gene “on” or
“off.” It is the portion of the
gene where cellular machinery
binds before transcribing the
DNA blueprint into a useful RNA
.
PROMOTER
5. In genetics, Promoters are DNA
sequences located in the 5' region
adjacent to the transcriptional start site.
RNA polymerase and accessory proteins
(transcription factors) bind to the
promoter to initiate production of an
mRNA transcript. Interactions of proteins
at the promoter regulate gene activity by
activating or repressing transcription
6. There is virtually an endless number of
promoters, potentially as many as there are
genes. Promoters, like genes, are made up
of A's, G's, C's and T's all lined up in a
certain order. Promoters can be about 100–
1000 base pairs long.The promoter region
can be short or quite long; the longer the
promoter is, the more available space for
proteins to bind.
7. Role in transcription
• The whole process of transcription starts
when the RNA polymerase binds to the
promoter. RNA Polymerase has 4 subunits
i-e alpha , beta , beta’ and sigma. Only the
Sigma Factor is required for RNA
polymerase to bind to the promoter. As the
RNA polymerase binds to the promoter ,
DNA duplex becomes unwind , base pairs
are broken down and transcription bubble
is appeared. This is followed by elongation
phase where the RNA grows and protrudes
from the bubble.
8.
9. TYPES OF PROMOTERS USED
FOR GENE EXPRESSION
1: Constitutive promoters :
●Induce the expression of the downstream located coding
region in all tissues irrespective of environmental or
developmental factors.
Example:
Plant pathogen promoters CaMV 35S promoter.
10. ●Operate in particular tissues and at certain
developmental stages.
●Maybe induced by endogenous and exogenous
factors.
Examples:
Tomato pz7 and pz10 gene promoters ( for ovary
gene expression)
2: Tissue Specific Promoters:
11. 3 : Inducible promoters:
● Within this group, there are promoters
modulated by presence or absence of biotic or
abiotic factors such as light, oxygen levels,
heat, cold and wounding.
Inducible promoters are grouped as:
a ) Chemically regulated promoters
b) Physically regulated promoters
12. 4 : Synthetic promoters :
● Promoters made by bringing together the
primary elements of a promoter region from
diverse origins.
Example:
Maize ubiquitin 1 gene ( Ubi 1) core promoter
Cytomegalovirus (CMV) promoter
CAG promoter
13. Promoter regions
There are three main portions that make up a
promoter: core promoter, proximal promoter, and
distal promoter.
CORE PROMOTER REGION:
The core promoter region is located most
proximally and contains the RNA polymerase binding
site, TATA box, and transcription start site (TSS).
RNA polymerase will bind to this core promoter
region stably and transcription of the template
strand will initiate.
14. PROXIMAL PROMOTER REGION:
Proximal promoter are upstream from the core
promoter which contain many primary regulatory
elements. The proximal promoter is found
approximately 250 base pairs upstream from the
TSS and it is the site where general transcription
factors bind.
15. DISTAL PROMOTER REGION:
The final portion of the promoter region is called
the distal promoter which is anything further
upstream from the gene. The distal promoter
also contains transcription factor binding sites,
but mostly contains regulatory element
16. PROKARYOTIC PROMOTERS
●Promoters in prokaryotic organisms are two short DNA
sequences located at the -10 (10bp 5' or upstream) and -35
positions from the transcription start site (TSS).
●Their equivalent to the eukaryotic TATA box, the
Pribnow box (TATAAT) is located at the -10 position and
is essential for transcription initiation.
● The -35 position, simply titled the -35 element, typically
consists of the sequence TTGACA and this element
controls the rate of transcription.
● Prokaryotic cells contain sigma factors which assist the
RNA polymerase in binding to the promoter region. Each
sigma factor recognizes different core promoter
sequences.
18. EUKARYOTIC PROMOTERS
• Eukaryotic Promoters are much more
complex and diverse than prokaryotic
promoters. Eukaryotic promoters span a
wide range of DNA sequence. There are
two parts :
●The core promoter or the basal promoter
● Upstream promoter element
Core promoter is constituted by TATA box and
transcriptional start site.
Initiation complex binds to the core promoter
upstream elements are responsible of the
regulation of transcription.
21. BIDIRECTIONAL
PROMOTERS
• Pairs of genes control by same promoter but located on
opposite strand and opposite direction. Their TSS are
separated by less than 1,000 bp.
• In general, they are rich in CpG content .
• Function of genes represented in bidirectional class are
often: DNA repair genes, chaperone protein, and
mitochondrial genes.
• Genes control by bidirectional promoters are often co-
express, but a minority of bidirectional genes have a
mutual exclusive expression.
• No correlation between length of promoter and degree of
expression
22. ●Bi-directional promoters are a common feature of
mammalian genomes About 11% of human genes are
bi-directionally paired.
●Certain sequence characteristics have been
observed in bidirectional promoters:
1: A lack of TATA boxes
2:An abundance of CpG islands
3:Symmetry around the midpoint of dominant Cs and
As on one side and Gs and Ts on the other.
23. Sub-genomic promoters
A sub-genomic promoter is a
promoter added to a virus for a
specific heterologous gene,
resulting in the formation of
mRNA for that gene alone.
24. A wide variety of algorithms have been
developed to facilitate detection of
promoters in genomic sequence, and
promoter prediction is a common element of
many gene prediction methods. A promoter
region is located before the -35 and -10
Consensus sequences. The closer the
promoter region is to the consensus
sequences the more often transcription of
that gene will take place.
Detection of
promoters
25. COMMONLY USED
PROMOTERS
The most commonly used promoters are:
Lac (Lactose )
Trp, ( Tryptophan )
Tac, ( invitro combo of Lac and Trp)
T 7 System
All of these bind to the sigma-70 factor
for transcription.
26. THE AMAZING T7
SYSTEM
The T7 system is very useful because T7 RNA
polymerase is an incredibly fast and powerful enzyme. We
can place the gene for T7 RNA polymerase behind the lac
promoter/operator, in a cell with lac Iq for tight regulation
of this protein; then, we can put our target genes behind T7
promoters. When we want the genes expressed, we flood
the cell with IPTG, which will induce the production of T7
RNA polymerase, which will, in turn, make massive numbers
of copies of the target genes located behind the T7
promoters.
27. OPERONS
Prokaryotes contain operons while eukaryotes do
not. Operons are clusters of related genes
involved in a similar function and are often found
in a contiguous array. Operons are controlled by a
single promoter.
Inducible Operons
An inducible operon is an operon where a
substance is required to be bound before
transcription will occur, it is normally "off" but
when a substance binds it is turned "on.“
Example: The lac operon .
28. Repressible Operon
A repressible operon is an operon that where
transcription is normally "on," but when a
substance binds it turns "off." It is the
opposite of an inducible operon.
Example:
The Trp Operon.
29. ●A promoter is classified as strong or weak according
to its affinity for RNA polymerase (and/or sigma
factor)
●This is related to how closely the promoter sequence
resembles the ideal consensus sequence for the
polymerase
Strong promoters will keep gene expression tuned to
the highest level with efficient gene expression.
STRENGTH OF A
PROMOTER
30. Diseases associated with
aberrant promoter
This is a list of diseases where evidence suggests
some promoter malfunction, through either direct
mutation of a promoter sequence or mutation in a
transcription factor or transcriptional co-activator.
Example:
Diabetes
Asthma
Beta thalassemia
Rubinstein-Taybi syndrome
31. Why the interest in
promoters
The interest in promoters stems from the myriad
opportunities for controlling gene expression. The
study and understanding of the function of their
multiple components and the factors associated with
their performance have opened up the possibility of
modulation of the expression of genes in
homologous organisms as well as in heterologous
organisms, where foreign promoters together with
genes of interest are inserted. Promoters are
regarded as molecular biological tools crucial for
the regulation of the expression of genes of interest.
As such, they have a huge influence in follow-on
research and development in biotechnology.