DNA Structu re

And
Repl icati on

Vasudevan 1
Bases- Purines
Pyrimidines

Purines : Adenine, Guanine

Pyrimidines : Thymine, Cytosine

Vasudevan 2
 OH
5' phosphate
_ end
O P O

H O-H2C Thymine
O

OH

OH
O H

O 3’P
_ 2’
O

O-H2C Cystosine
O

OH

O H
Base + Deoxy ribose
O P O
_ = Nucleoside
Adenine
O-H2C O

OH
Vasudevan OH H 3
3' OH end
 OH
5' phosphate
_ end
O P O

O-H2C Thymine
O

OH

OH
O H

O 3’P
_
O

O-H2C Cystosine
O

OH

O H
Base + Deoxy riboseO+ Phosphoric
_ acid
P O
Adenine
O-H2C O = Nucleotide
OH
Vasudevan OH H 4
3' OH end
 OH
5' phosphate
_ end
O P O

O-H2C Thymine
O

OH

O H
Phospho _
Diester O P O

linkage O-H2C O
Cystosine

OH

OHO
_
H

O3’P O
Adenine
O-H2C O

OH
Vasudevan OH H 5
3' OH end
 OH
5' phosphate
_ end
O P O

O-H2C Thymine
O

OH

O H
Phospho _
Diester O P O

linkage O-H2C O
Cystosine

OH

O H
_
O P O
5’-T-C-A-3’ O-H2C O
Adenine

Polarity
OH
Vasudevan OH H 6
3' OH end
5’ end 3’ end

Phosphate
bonds

Double
Helix
Bases
jutting
inside
Vasudevan 7
 Hydr ogen
bonds
between
bases

Base pairing rule

A to T
G to C

Nucl eoti dVasudevan 8

Watson
1962 Edwin
Chargaff
1905- 2002
Crick
1962

Wilkins
Rosalind
1962
Franklin
Vasudevan
1921-1958 9
 Watson-Crick Model of DNA
1. Right handed Double Helix
2. Base pairing rule, A toT; G to C
by Hydrogen bonding
3. Antiparallel 5’ 3’
3’ 5’
4. 10 base pairs form one spiral

Vasudevan 10
 Cor e histones as octamer

DN A strand

DNA wraps twice around

histone octamer to form
one nucleosome
Vasudevan 11
5 Finally condensed as chromosomes

4 Condensed to
chromatin

3 Forms Supercoil

2 Forms
Nucleosomes

1 DNA double helix

Vasudevan 12
5 Finally condensed as chromosomes

4 Condensed to
chromatin

3 Forms Supercoil

2 Forms
Nucleosomes

1 DNA double helix

Vasudevan 13
5 Finally condensed as chromosomes

4 Condensed to
chromatin

3 Forms Supercoil

2 Forms
Nucleosomes

1 DNA double helix

Vasudevan 14
5 Finally condensed as chromosomes

4 Condensed to
chromatin

3 Forms Supercoil

2 Forms
Nucleosomes

1 DNA double helix

Vasudevan 15
5 Finally condensed as chromosomes

4 Condensed to
chromatin

3 Forms Supercoil

2 Forms
Nucleosomes

1 DNA double helix

Vasudevan 16
 Inactiv ation of D NA
During
Dif ferenti ation
All human cells are derived from a
single cell, the zygote. Therefore,
all cells contain the same genetic
information.

In a cell, about 90% DNA are

permanently inactive.

Diff erentiat ion .

Vasudevan 17
 Intr ons, Exons, C istr ons

Only about 10% of the human DNA

contains genes; the rest silent
areas.
The segments of the gene coding
for proteins are called exons
(expressed regions).

They are interspaced in the DNA

with stretches of silent areas,
called introns (intervening areas).
Vasudevan 18
The primary transcripts contain
intron sequences;

which are later removed to

produce mature mRNA.

Introns are not translated.

CISTRONS
Vasudevan 19
 REPLICATION
Synthesis of new DNA strand
on the basis of template strand

Semiconservative
replication
A new complementary strand is
synthesised over the old template

Vasudevan 20
 Parent cell
DNA

First
generation

Second
generation

Meselson (Left)
and Stahl (Right)
Vasudevan 21
 Conservative Semi -
replication conservati ve
(theoretical; repl icati on
but actually (actual )
not taking place)

Vasudevan 22
 Conservative Semi -
replication conservati ve
(theoretical; repl icati on
but actually (actual )
not taking place)

Vasudevan 23
Old strands

Parent strands
Partially
unwound

New strands made

Based on
Base Pairing rule

Old New Old

Vasudevan
New 24
 Salient features of Replication

1.Each strand serves as a TEMPLATE

over which new COMPLEMENTARY
strand is synthesised

2. Base parining rule, A with T; G to C

3. Polymerisation of the new

strand is taking place from 5’ to 3’
direction
Vasudevan 25
 DNAPolymerase (DNAP)
5 in number; alpha is main enzyme

OH

5’ 3’
Arthur Kornberg
NP 1959
Vasudevan 26
 DNAPolymerase action

OH

5’ 3’

Vasudevan 27
 DNAPolymerase action

OH

5’ 3’

Vasudevan 28
 DNAPolymerase action

5’ 3’

Vasudevan 29
 RNA primer is needed for DNA synthesis

RNA primer primase

Old DNA strand

DNA polymerase

Another DNAP New DNA strand

Completed new DNA strand

Vasudevan 30
RNA primer primase

Old DNA strand

RNA primer DNA polymerase

Another DNAP New DNA strand

Completed new DNA strand

Vasudevan 31
 Old DNA strand
RNA primer DNA polymerase

Another DNAP New DNA strand

Completed new DNA strand

Vasudevan 32
 RNA primer is needed for DNA synthesis

Old DNA strand

DNA polymerase

Another DNAP New DNA strand

Completed new DNA strand

Vasudevan 33
Replication bubble (Replication fork)

3’ 5’

3’
5’

New DNA
DNA uncoils RNA primer

Vasudevan 34
 Lagging strand and Okazaki pieces
3’
5’

5’ Old strand Leadi ng s trand

3’
Short RNA pr imer
3’ Di recti o Okazaki fragment
n
of fork Poi nt of joini ng
movem e 3’ af ter
nt
5’ removi ng RN A prmer
Laggi ng st rand
Vasudevan 35
 Lagging strand and Okazaki pieces
3’
5’

5’ Old strand Leadi ng s trand

3’
Short RNA pr imer
3’ Di recti o Okazaki fragment
n
of fork Poi nt of joini ng
movem e 3’ af ter
nt
5’ removi ng RN A prmer
Laggi ng st rand
Vasudevan 36
 Lagging strand and Okazaki pieces
3’
5’

5’ Old strand Leadi ng s trand

3’
5’
Short RNA pr imer
3’ Okazaki fragment
3’ Di recti o
n
of fork Poi nt of joini ng
movem e 3’ af ter
nt
5’ removi ng RN A prmer
Laggi ng st rand
Vasudevan 37
 Lagging strand and Okazaki pieces
3’
5’

5’ Old strand Leadi ng s trand

3’
5’
Short RNA pr imer
3’ Di recti o Okazaki fragment
n Or
of fork 3’ Poi nt of joini ng
movem e Okazaki
3’ af ter pieces
nt
5’ removi ng RN A prmer
Laggi ng st rand
Vasudevan 38
 Lagging strand and Okazaki pieces
3’
5’

5’ Old strand Leadi ng s trand

3’
5’
Short RNA pr imer
3’ Di recti o Okazaki fragment
n
of fork Poi nt of joini ng
movem e 3’ af ter
nt
5’ removi ng RN A pri mer
Laggi ng st rand
Vasudevan 39
 Lagging strand and Okazaki pieces
3’
5’

5’ Old strand Leadi ng s trand

3’
5’
Short RNA pr imer
3’ Di recti o Okazaki fragment
n
of fork Poi nt of joini ng
movem e 3’ af ter
nt
5’ removi ng RN A prmer
Laggi ng st rand
Vasudevan 40
 Summary of DNA replication
1. Unwinding of parental DNA to
form a replication fork.
2. RNA primer complementary to
the DNA template is synthesised
by RNA primase.
3. DNA synthesis is continuous in
leading strand (towards replica- tion
fork) by DNA polymerase.
4. DNA synthesis is discontinuous in
lagging strand (away from the
fork), as Okazaki Vasudevan
fragments. 41
 Summary of DNA replication
5. In both strands, the synthesis is
from 5' to 3' direction.

6. Then the RNA pieces are removed;

the gaps filled by deoxynucleo-
tides and the pieces are ligated by
DNA ligase.
7. Proof reading is done by the DNA
polymerase.

8. Finally organised
Vasudevan
into chromatin.
42
 Old template strand

contains
methylated

bases; so wrong
base could be
identified

NUCLEOTIDE EXCISION
REPAIR (NER) MECHANISM
Vasudevan 43
 Old template strand

contains methylated

A portion
bases; so wrong base
could
aroundbe identified

that area is cut

and removed by
endonuclease

NUCLEOTIDE EXCISION
REPAIR (NER) MECHANISM
Vasudevan 44
 Old template strand

contains methylated

bases; so wrong base

A portion
could around
be identified
that area is cut
and removed by
endonuclease

Gap is filled with

correct base
sequences by
DNA polymerase
NUCLEOTIDE EXCISION
REPAIR (NER) MECHANISM
Vasudevan 45
 Old template strand

contains methylated

bases; so wrong base

A portion
could around
be identified
that area is cut and
removed by
endonuclease
Gap is filled with
correct base
sequences by DNA
polymerase
Finally DNA
Vasudevan
ligase 46
seals the nicks
Diseases associated with
DNA repair mechanisms
1. Xeroderma Pigm en tosum :
Defective NER mechanism;
sensitivity to UV light; skin cancers
2. Ataxi a Telangectasi a :
defective ATM gene; sensitivy to UV
light; lymphoreticular neoplasms
3. Fanconi 's Anemi a : Defect
in DNA repair; increased occurrence
of cancer Vasudevan 47
 DN AP coul d not r epl icate thi s area
(Tel omer e)
3'
5'
New tel omere repeat
Tel omerase

RN A templ ate
ins ide
tel omer as e

DN A pol ymerase
RN A produced by
compl etes
Vasudevantel omer as e acts as
laggi ng str and 48
pri mer
 DN AP coul d not r epl icate thi s area
(Tel omer e)
3'
5'
New tel omere repeat
Tel omerase

RN A templ ate
ins ide
tel omer as e

DN A pol ymerase
RN A produced by
compl etes
Vasudevantel omer as e acts as
laggi ng str and 49
pri mer
DN AP coul d not r epl icate thi s area
(Tel omer e)
3'
5'
New tel omere repeat
Tel omerase

RN A templ ate
ins ide
tel omer as e

DN A pol ymerase
RN A produced by
compl etes
Vasudevantel omer as e acts as
laggi ng str and 50
pri mer
Inhibitors of DNA replication

1. Anti bacterial agents

a)Ciprofloxacin
Inhibits Bacterial DNA gyrase

b)Nalidixic acid do

c)Novobiocin do
Vasudevan 51
2. An ti cancer agents

a)Etoposide Human
topo-isomerase
b)Adriamycin do
c)Doxorubicin do
d)6-mercaptopurine Human
DNA polymerase
e)5-fluoro uracil do

Vasudevan 52
What enz ym es are requi red f or
DN A repl icati on?

DN A pol ymerase
Topo iso mer ase
DN A ligase

Vasudevan 53
Repl ic at ion is in whi ch di rect ion?

Pol ymeri sat ion of the new strand

of DN A is f rom 5’ to 3’ di recti on

What is semi -conservative

natur e of repl icati on?
In the daughter c ell , one strand
is de rived from the mother cel l;
whi le the other strand is n ewly
synthesi sed
Vasudevan 54
What is semi -di sconti nuou s
natur e of repl icati on?

In the l eadi ng str and, the

repl ication is co ntinuous;
but in the laggin g strand,
repl ication is taki ng pl ace
in smal l pi eces.

Vasudevan 55
What is l aggi ng str and?

The stran d i n whi ch rep licati on

is dis- conti nuous is cal led
Laggi ng st rand

What are O kaz aki fragments?

The saml l DN A m ole cul es
attached to its own pri mer RN A
in the laggi ng st rand are call ed
Okazaki piec es.
Vasudevan 56
Name the drug s wh ich inhi bi t
DN A repl icati on in prokaryotes
Ci prof loxaci ne, Novobi oci n

Name some anti cancer drugs

whi ch in hibi t DN A re pl icati on i n
ammal ian cel ls
5- fluorouraci l
6- mercaptopurine
Cytosi ne a rabi nosi de
Etoposi de Vasudevan 57