Molecular Biology

DNA structure


amino acid
Regions oI hydrogen bonding
between base pairs

enome
W %e total number of genes in one cell constitutes
te genome of te cell.
W %ere are about 3 X 10
9
base pairs (bp) in eac
uman aploid genome. %ey are distributed in
23 cromosomes.
W nly 10% of te total uman genes are
functioning genes.
W ene:
t is a deoxyribonucleic acid (DNA)
segment, or in some viruses a ribonucleic
acid (RNA) segment tat contains a well
defined genetic information.
W %e genes are carried on cromosomes.
W Because cromosomes are paired,
everybody carries two copies of eac
gene.
Replication
Transcription
mRNA
DNA
protein
Translation
ene Expression
DNA Replication
W Definition:
t is te process of DNA syntesis using
parent DNA strands as template.
W t aims at formation of a copy of te parent
DNA molecule for te daugter cell
$tages of DNA replication in
eukaryotes
- nitiation
- Elongation
- %ermination
nitiation of DNA replication
1- dentification of te origins of replication
2- Unwinding of double stranded DNA
3- Formation of replication forks
4- RNA primer syntesis
/entification of the origin of repIication
ormation of repIication fork
ormation of RepIication fork
&nwin/ing of /oubIe stran/e/ DNA
Elongation of DNA replication
1- Leading strand syntesis
2- Lagging strand syntesis
%ermination of DNA
replication
Rules of DNA replication in
eukaryotes
1- DNA replication is semiconservative
2- Replication begins at multiple origins and
usually bidirectionally
3- Replication exibits polarity
4- Replication is very accurate
5- DNA replication occurs in te nucleus
during te syntetic pase of eukaryotic
cell cycle
Basic requirements for DNA
replication
1- $ubstrates: %e four deoxynucleotide
tripospates
2- %emplate
3- Primer: DNA polymerases are unable to
start DNA syntesis except in te
presence of pre-existing primer
4- Enzymes: elicase, topoisomerases, DNA
polymerases and ligases
DNA Repair
W t is a mecanism to repair damaged DNA
W DNA is subject to damage by a wide
variety of agents. %erefore, te presence
of a mecanism of DNA repair to maintain
te genetic information is very important.
W f te damage is not repaired, a permanent
mutation may be introduced tat can result
in serious effects.
auses of DNA damage
1- $pontaneous canges:- e.g. *Deamination of
cytosine to uracil.
* Depurination.
2- DNA polymerase errors:- e.g. 2-amino purine
incorporated instead of adenine into a newly
syntesized DNA strand.
3- radiation damage: * U.V.
* onizing radiation (X-ray, -rays).
4- emicals:
* Nitrous acid ( HN2).
* Alkylating agents such as dimethyl
sulfate
5- xidative damage by: * H22.
* Hydroxyl radicals.
6- arcinogenic compounds in food, water or air
e.g. * Aflatoxin
* emical (in cigarette smoke).
%ypes of DNA Repair
1- Mismatc repair.
2- Base excision repair.
3- Nucleotide excision repair.
4- Double strand break repair
Defects in DNA Repair ause
Human Diseases
eroderma Pigmentosa:
W t is an autosomal recessive disease.
W %e most common deficiency occurs in te
excision endonuclease enzyme.
W t is caracterised by extreme sensitivity to
sunligt, ulceration and skin cancer
%ranscription
W t is te process of syntesis of RNA from
a template by RNA polymerase
enzymes and a number of associated
proteins
Basic features of RNA syntesis
1- %e precursors of RNA syntesis are te
four ribonucleotide 5`- tripospates.
2- %e RNA cain grows in te 5` 3`
direction wic is te same as te
direction of cain growt in DNA syntesis
3- RNA polymerases, in contrast with DNA
polymerases, can initiate RNA syntesis ,
no primer is needed
4- ontrary to DNA replication, only one
strand is transcribed, unwinding is limited
to te transcribed gene segment
$teps of RNA syntesis
1- nitiation
2- Elongation
3- %ermination
omparison of DNA synthesis (repIication) an/ RNA synthesis (transcription)
RepIication Transcription
* Definition DNA synthesis from DNA tempIate. RNA synthesis from DNA
tempIate.
* haracter ompIete
(The entire chromosome is
normaIIy copie/).
SeIective
(OnIy particuIar genes are
transcribe/ at any one time,
an/ some portions of the DNA
genome are never
transcribe/).
* TempIate The two DNA stran/s serve as
tempIate.
OnIy one of the two stran/s
serves as tempIate.
* PoIymerization DNA poIymerase enzyme
(DNA poIymerase cannot initiate
stran/ synthesis).
RNA poIymerase.
(RNA poIymerase can).
- RNA primer Require/ Not require/
- Require/
substrates
/ ATP, / TP,
/ TP an/ / TTP
ATP, TP,
TP, &TP
- Direction of
synthesis
5` 3` 5` 3`
- proofrea/ing
activity (3`
5` exonucIease).
Present
(high - fi/eIity DNA synthesis).
Absent
(Iow- fi/eIity RNA synthesis).
Post- transcriptional modification of
RNA
Processing of eukaryotic mRNA
precursor:
1- apping
2- Polyadenylation
3- $plicing
(1) apping:-
W %e cap (7-metyl guanosine) is added to te 5` end
wile te RNA molecule is still being syntesized.
W 7- metyl guanosine is joined to te 5` end of mRNA in
an unusual 5`, 5` - tripospate linkage.
$31.a3.e o1 the .ap stru.ture:-
W $erves as a ribosome binding site (for translation
initiation).
W Protects te 5` end of mRNA from attack by 5` 3`
exonuclease (for mRNA stability).
(2) PoIya/enyIation:-
W Poly (A) polymerase enzyme adds poly (A) tail to
te 3`end of mRNA (wic is sort at first about
20A residues , ten subsequently extended to as
muc as 200 A residues).
$31.a3.e o1 the poly ( tal:-
W Protects te 3` end of mRNA from 3` 5`
exo3u.lease attack (for mRNA stability).
W Aids in its transport from nucleus to cytoplasm.
(3) SpIicing:-
$plicing means removal of introns from te
primary transcript in te nucleus and ten
ligation of exons to form mature functional
m RNA. %e mature mRNA is ten
transported to te cytoplasm were it is
translated into protein
Processing of eukaryotic ribosomaI RNA
precursor (pre- r RNA):-
W Eukaryotic rRNA is transcribed in te nucleolus
by # polymerase I as a single piece of 45 $
RNA precursor (pre- rRNA)
W %en it is subsequently cleaved to yield 28 $
rRNA, 18 $ rRNA, and 5.8 $ rRNA.
W RNA polymerase transcribes te 5$ rRNA unit
from a separate gene.
Processing of eukaryotic transfer RNA
precursor (pre- tRNA):-
W Eukaryotic tRNA genes are all transcribed by
RNA polymerase .
W %e primary transcript (pre- tRNA) requires post-
transcriptional processing to be mature
functional tRNA suc as:-
- Folding and base pairing to generate its
caracteristic sape.
- Removal of excess nucleotides from te 5`
and 3` ends (cleavage).
- Removal of introns (splicing).
- Addition of te A sequence at 3`
end.
- Modification of some bases by
metylation, deamination or reduction.