DNA REPLICATION

 Discoveries of DNA structure (Watson &

Crick, 1953), solve the mistery.

1.How does DNA carry information.

2.How this information copied before cell

devide.
 DNA REPLICATION

BUT THE MANNER IN WHICH

REPLICATION OCCURRED
REMAIN A MYSTERY
 DNA REPLICATION
 TREE POSSIBLE MODELS
1.Conservative Model
2.Semiconservative Model
3.Dispersive Model
 Distinguish
 Grew bacteria with N15(heavy Nitrogen)
for 1 & 2 round of cell division with the
presence of N14(normal Nitrogen).
 Harvested and isolated (centrifugation)
 1st round only one band formed at the
midle (between N15 & N14 band) – So
reject Conservative model
 2nd round two bands formed only for
Semiconservative model.
 5
DNA
O 3

3 O
P 5 P
5 O
1 G C 3
2
4 4
2 1
3 5
P O
P
5
T A 3
O

O
5
P 3 P
 DNA REPLICATION
 Catalyzed by a team of enzyms
1.Unwind double helix into two portion of
templates strand at origin of replication.
– DNA helicase enzyme break hydrogen bond.
2.Preventing re-formation of double helix
– helix-destabilizing proteins by binding to single
DNA,
- topoisomerase produced breaks in the
DNA molecules & then rejoin the strands,
relieving strain and effectively preventing the
formation of knots during replication.
DNA REPLICATION
 - The regions of active DNA synthesis is
associated with replication fork (junction
of the single strands and the double-
stranded region – Y shape.
3. DNA polymerase attach free nucleotide to
template strand in the vacinity of the fork
and reformation phophdiaster bond.
Anergy suplied from broken 2 molecule
phosphate of ATP (free nucleotide).
4. Adding new nucleotides only to the 3’ end
- linkage of the 5’ phosphate group of the
next nucleotide subunit to the end 3’
hydroxyl group of the sugar, so new strand
always grows in the 5’ 3’ direction.
5. Adding new nucleotides only to the
existing polynucleotide strand.
6. RNA primer (5-14 nucleotides) is first synthesized
at the point of initiotion of replication (by primase
enzyme).
7. Since two template strands are antiparallel 3’ 5’
there are two identical DNA polymerase move
opposite direction during replication.
8. New strand smooth and continuous move toward
replication fork is called leading strand.
9. Opppsite direction is called laging strand, only
short pieces can be synthesized (Okazakie
fragments, 100-200) since DNA polymerase
cannot away from the fork.
10.Each OF initiated by separate RNA
primer and move towards 5’end of
previously synthesized fragment.
11. When the two fragment are reached,
both will then joint together:
 DNA Pol I (exonuclease) -removes RNA Primers.
 The gaps are closed with the action of DNA Polymerase
(adds complementary nucleotides to the gaps)
 DNA Ligase (adds phosphate in the remaining gaps of
the phosphate - sugar backbone).
 The DNA Polymerase reaches to an end of the
strands.
 The DNA Polymerase seal the gap (because
there is no RNA primer).
 So, the end of the parental strand where the last
primer binds isn't replicated.
 These ends of linear (chromosomal) DNA
consists of noncoding DNA that contains repeat
sequences and are called telomeres.
 A part of the telomere is removed in every cycle
of DNA Replication.
 Fixes possible errors coused during the
replication
 Enzymes like nucleases remove the
wrong nucleotides and the DNA
Polymerase fills the gaps.
 Proofreading New DNA
 DNA polymerase initially makes
about 1 in 10,000 base pairing
errors
 Enzymes proofread and correct
these mistakes
 The new error rate for DNA that
has been proofread is 1 in 1 billion
base pairing errors
 DNA Damage & Repair
 Chemicals & ultraviolet radiation
damage the DNA in our body cells
 Cells must continuously repair
DAMAGED DNA
 Excision repair occurs when any of
over 50 repair enzymes remove
damaged parts of DNA
 DNA polymerase and DNA ligase
replace and bond the new nucleotides
together
 Question:
 What would be the
complementary DNA
strand for the following
DNA sequence?

DNA 5’-CGTATG-3’
 Answer:

DNA 5’-GCGTATG-3’
DNA 3’-CGCATAC-5’