REPLIKASI

DNA

Dr. Oeke Yunita, S.Si., M.Si., Apt.

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Synthesis Phase (S phase)

S phase during interphase of the
cell cycle
Nucleus of eukaryotes

phase

DNA replication takes

place in the S phase.

G1

interphase

G2

Mitosis

-prophase
-metaphase
-anaphase
-telophase
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DNA Nucleotide
Phosphate
Group

O=P-O
O

CH2

O
N
C1

C4

Nitrogenous base
(A, G, C, or T)

Sugar
(deoxyribose)

C3

C2

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TAHAP TAHAP
REPLIKASI DNA
1. Denaturasi (Pemisahan) untaian DNA
template
2. Inisiasi
3. Pemanjangan untaian DNA
4. Ligasi fragmen DNA
5. Terminasi

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DNA Replication
Begins at Origins of Replication
Two strands open forming Replication
Forks (Y-shaped region)
New strands grow at the forks
3

Parental DNA Molecule

Replication
Fork

3
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DNA replication system

Template:

double stranded DNA

Substrate:

dNTP

Primer:

short RNA fragment with a

free 3-OH end

Enzyme:

DNA-dependent DNA
polymerase (DDDP),
other enzymes,
protein factor
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Players involved in DNA Replication

Origin of Replication Enzymes (Initiator Proteins) recognize the start site on

template DNA and initially open up the DNA (single strands)

Helicases untwist the double helix at the replication forks

Primase begins DNA replication by laying an an RNA primer (10 nt long) for DNA
polymerase to start at

Single-strand binding proteins bind to and stabilize the single-stranded DNA

until it can be used as a template

DNA polymerase polymerizes new DNA in a 5 to 3 direction, proofreads the

newly laden DNA, excises error ridden DNA and replaces that DNA with the
correct sequence. Speed/fidelity: mutates 1 X 10-10

Topoisomerase corrects overwinding ahead of replication forks by breaking,

swiveling, and rejoining DNA strands

Terminal replication proteins stall replication fork progress for polymeriation

to catch-up

DNA ligase anneals nucleotides together, creating a DNA strand

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III

III

I
DNA

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Replication of Strands
Point of Origin

Replication
Fork

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Components of the DNA Replication

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Core proteins at the replication fork

Nature (2003) vol 421,pp431-435

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DNA Primer synthesis

On Lagging strand

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DENATURASI
Initiating Proteins for DNA replication
1. Initiator protein, 2. helicase binding
to initiator protein, 3. helicase loading
on DNA, 4. helicase opens the DNA and
binds to primase, 5. RNA primer
synthesis, 6. DNA polymerase binding
and DNA synthesis

INISIASI
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Replication
PROKARYOTE
Replication is bidirectional from a
specific sequence of DNA called the
origin of replication (OriC)
DnaA (Initiator protein) binds to
the 9 bp repeat sequence within
OriC, causing local denaturation of
the helix within the 13 bp repeat
DNA helicase is recruited (by DnaA)
and loaded (by DnaC), and untwists
the DNA helix (use ATP)
Helicase also recruits DNA primase
(modified RNA pol), forming the
primosome complex, which makes
an RNA primer of 5 -10 nucleotides)
for DNA synthesis
Helical tension is relieved by DNA
gyrase (topoisomerase)
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Synthesis of telomeric DNA by telomerase

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Step 1 = Binding

The bindingpolymerizationtranslocation cycle can

occurs many times

Step 2 = Polymerization

This greatly lengthens

one of the strands

Step 3 = Translocation

The complementary
strand is made by primase, DNA
polymerase and ligase

RNA primer
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II. The Eukaryotic Problem of Telomere Replication

RNA primer near end of
the chromosome on
lagging strand cant be
replaced with DNA since
DNA polymerase must add
to a primer sequence.

Do chromosomes get shorter with

each replication???
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Solution to Problem: Telomerase

Telomerase enzyme adds TTGGGG
repeats to end of lagging strand
template.

Forms hairpin turn primer with free

3-OH end on lagging strand that
polymerase can extend from; it is later
removed.

Age-dependent decline in telomere

length in somatic cells, not in stem
cells, cancer cells.

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mtDNA replication

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