History

R strain could become

Experimen dg Streptococcus pneumoniae
virulent when it took in
galur :
DNA from heat-killed S
Smooth (S) – Virulent (gel coat)
strain
Rough (R) – Kurang Virulen
Structure
DNA Nucleotide
Phosphate
Group

O 5
O=P-O CH2
O
O
N
Nitrogenous base
C4 C1 (A, G, C, or T)

Sugar
(deoxyribose)

C3 C2
 Deoksi adenosin monofosfat

Deoksi guanosin monofosfat

 Deoksi timidin monofosfat

Deoksi sitidin monofosfat

 Melting and Renaturation of DNA

Renaturation driven by homologous base pairing

Will also hybridize with a radiolabeled 5’-ACGGCTA-3’ “probe”.

 Senyawa yang menstabilkan kondisi terdenaturasi

O O

NH2  C  NH2 NH2  C  H

Urea Formamid
Replication
 Process of duplication of the entire
genome prior to cell division
 In eukaryotes , replication only occurs
during the S phase of the cell cycle.
Synthesis Phase (S phase)
• S phase during interphase of the
cell cycle
• Nucleus of eukaryotes
S
DNA replication takes phase
place in the S phase.

G1 interphase G2

Mitosis
-prophase
-metaphase
-anaphase
-telophase
DNA replication occurs with great fidelity
(New cells will need identical DNA strands))

Somatic cell DNA stability and reproductive-cell DNA stability are essential.
Why?

Identity

Pan troglodytes
98.77% sequence identity

Genetic diseases
 Basic rules of
replication

A. Semi-conservative
B. Starts at the ‘origin’
C. Bidirectional
D. Semi-discontinuous
E. Synthesis always in the 5’-3’ direction
F. RNA primers required
DNA replication
Of the 3
possible
models, 
replication
is…

A) Semi-
conservative

Meselson-Stahl
experiments
 B) Starts at origin
Initiator proteins identify specific base
sequences on DNA called sites of origin

Prokaryotes – single origin site E.g E.coli - oriC

Eukaryotes – multiple sites of origin (replicator)
E.g. yeast - ARS (autonomously replicating
sequences)

Prokaryotes Eukaryotes
Bidirectional replication of circular
DNA molecules.
Temporal ordering of DNA replication initiation events in replication units of
eukaryotic chromosomes.
 C) bidirectional
 Replication forks move in one or opposite directions


 D) Semi-discontinuous replication
Anti parallel strands replicated simultaneously
 Leading strand synthesis continuously in 5’– 3’

 Lagging strand synthesis in fragments in 5’-3’
E) Synthesis is ALWAYS in the 5’-3’ direction
Nucleotide recognition
Enzyme catalysed polymerisation (DNA polymerase)
Complementary base pair copied
Substrate used is dNTP
 F) RNA primers required
• DNA polymerase can only join an incoming nucleotide to one that
is base-paired

• RNA primase provides a base paired 3’ end as a starting point for
DNA pol by synthesising ~10 nucleotide primers
Animasi replikasi
 exonuclease 3’-5’

exonuclease 5’-3’

SSB (ssDNA binding protein) Binds to and stabilizes ssDNA

What happens if a base Where does energy for addition
mismatch occurs? of nucleotide come from?

From cleavage of high energy phosphate

DNA polymerase has 3’  5’ of incoming triphosphate
exonuclease activity in order to
correct errors
Why does DNA replication only occur in the 5’ to 3’ direction?
 DNase I

DNase II
Exonuclease
Replication of the ends of linear DNA
Since all known DNA polymerases
need a primer, how are the ends of
linear DNA replicated in eukaryotes?
newly synthesized DNA
RNA primer

5' 3'
template
Telomeres
repetitive DNA at the end of linear
eukaryotic chromosomes
Example
(GGGGTT)n n = 20 - 200

GGGGTT GGGGTT GGGGTT

5'
Telomerases : enzymes that add DNA
repeats to the 3' end of DNA.

Telomerases are
composed of protein and
an RNA molecule that AACCCCAAC

functions as the template

telomerase
for telomere synthesis.
 Human telomerase

 Responsible for maintaining telomere length in

eukaryotic chromosomes
 Main components:
 Telomerase reverse transcriptase
 Human telomerase RNA (hTR)
 Reverse transcriptase
 Transcribes RNA to DNA (rather than the usual DNA
to RNA)
 hTR is the template for the repeated region
5'

AACCCCAAC
GGGGTTGGGGTT
5'
telomerase
 AACCCCAAC
GGGGTT GGGGTT GGGGTT
5'

primase

GGGGTT GGGGTT GGGGTT

5'
 pol III

5'
pol I

ligase

telomeric repeats
For most cells, telomeres are added
during development. Later
telomerase becomes inactive.

Hence, as cells divide the DNA

becomes shorter.

Note that telomerase is reactivated in

many types of cancer cells.
OBAT anti REPLIKASI DNA
INHIBITOR TOPOISOMERASE (Gyrase)

Antibiotik QUINOLON : MENGHAMBAT

TOPOISOMERASE BAKTERI GRAM NEGATIF,
MODIFIKASI BAKTERI GRAM POSITIF
DAN AEROBIK

Camptothecin : INHIBITOR TOPOISOMERASE I

SEBAGAI ANTI KANKER DENGAN
MENSTABILKAN BENTUK ENZIM TERIKAT
PADA DNA SECARA KOVALEN
 TOPOISOMERASE SBG TARGET
OBAT
 Novobiocin – subunit ATPase GyrB
 Asam naladiksat – Gyr A
 Ciprofloxacin (oral) – stop replikasi

MENGGANGU PROSES PEMOTONGAN DAN PENYAMBUNGAN

UNTAI DNA