Replikasi dan Siklus Sel

Evi Umayah Ulfa, M.Si., Apt

 Pendahuluan
• Bagaimana materi genetik itu digandakan
sehingga sel anakan memiliki informasi
genetik yang sama dengan induk?
• Bagaimana informasi genetik itu digunakan
untuk mensintesis protein?
 Pendahuluan: Dogma Genetik
• REPLIKASI DNA →
BIOSINTESIS DNA

• SINTESIS PROTEIN →
BIOSINTESIS RNA →
PROTEIN
 Sentral Dogma
(Arus informasi Genetik)
Replikasi

Reverse
transcription
mRNA
(virus)
 REPLIKASI DNA
• Unidirectional, hanya satu strand DNA yang
bereplikasi
• Bidirectional, kedua strand DNA bereplikasi
dengan arah yang saling berlawananan
• Origin of replication: sisi dimana replikasi DNA
dimulai
prokariota → 1 sisi ori
eukariota → multiple sites of ori
 REPLIKASI DNA
• Sintesis DNA dengan cetakan DNA
• Tahap replikasi DNA
1. Inisiasi
2. Elongasi
3. Terminasi
• DNA template: leading dan lagging strand
• Primer: yang mengawali polimerisasi DNA
• Arah replikasi selalu dari 5’→ 3’
 KOMPONEN REPLIKASI DNA
1. Situs ori : Origin of replication
2. DnaA : mengenali ori
3. Helikase, enzim yang membuka double strand DNA
3. Single strand DNA Binding Protein, menstabilkan DNA rantai
tunggal
4. Gyrase, melepaskan “perputaran” DNA sehingga
mempermudah dan menstabilkan pembukaan
5. RNA polymerase untuk membuat primer (Primase) untuk
menginisiasi polimerisasi dengan menyediakan 3’-OH untuk
DNA Polymerase III
6. DNA Polimerase III, I
7. DNA ligase : menyambung celah okazaki
 REPLIKASI DNA: Elongasi
• Perpanjangan basa nukleotida dengan enzim
yang berperan utama: DNA Polymerase III
pada leading strand, DNA Polymerase I dan III
pada lagging strand (discontinues replication)
• Discontinues replication → terbentuknya
okazaki fragment pada lagging strand, o.k.
memerlukan DNA ligase untuk menyambung
fragmen-fragmen oligonukleotida
ARAH REPLIKASI
INGAT
DNA polimerase tidak bisa mengawali
sintesis dari nukleotida pertama
sehingga harus selalu ada „inisiator
nukleotida“

RNA Polimerase melakukan fungsi ini

dgn membentuk „primer“

Dan arah replikasi primer selalu dari

5‘ ke 3‘, sehingga hasilnya selalu ujung
3‘, disinlah baru DNA pol. berperan

Oleh karena itu terbentuklah leading

dan lagging strand yang tergantung
pada apakah primernya searah
dengan arah garpu replikasinya

Jika searah → leading strand

Jika tidak searah → lagging strand →

fragmen okazaki
 REPLIKASI DNA: Terminasi
• Pembuangan “RNA primers” dan pengisian gap oleh
DNA polymerase I

ERROR in REPLICATION
• DNA repair dan proofreading mechanism merupakan
mekanisme untuk mengatasi adanya error dalam
proses replikasi (maupun kerusakan DNA)
• Kerusakan dan kesalahan dalam replikasi ini jika tidak
diatasi akan menyebabkan mutasi yang berbahaya
Replikasi pd Eukariot
 Multiple Origins of Replication
 Eukaryotes have long linear chromosomes
 They therefore require multiple origins of replication
 To ensure that the DNA can be replicated in a reasonable time

 DNA replication proceeds bidirectionally from many

origins of replication
 Refer to Figure 11.20

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11-64
 Bidrectional DNA
synthesis

Replication
forks will
merge

Figure 11.20

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11-65
 Multiple Origins of Replication
 The origins of replication found in eukaryotes have some
similarities to those of bacteria

 Origins of replication in (Yeast)S. cerevisiae are termed ARS

elements (Autonomously Replicating Sequence)
 They are 100-150 bp in length
 They have a high percentage of A and T
 They have three or four copies of a specific sequence
 Similar to the bacterial DnaA boxes

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11-66
 Multiple Origins of Replication
 Origin recognition complex (ORC)
 A six-subunit complex that acts as the initiator of eukaryotic
DNA replication
 It appears to be found in all eukaryotes

 Requires ATP to bind ARS elements

 Single-stranded DNA stimulates ORC to hydrolyze ATP

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11-67
 Eukaryotes Contain Several
Different DNA Polymerases
 Mammalian cells contain well over a dozen different
DNA polymerases
 Refer to Table 11.4

 Four: alpha (a), delta (d), epsilon (e) and gamma (g)
have the primary function of replicating DNA
 a, d and e  Nuclear DNA
 g  Mitochondrial DNA

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11-68
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11-69
Nucleosomes and DNA Replication
 Replication doubles the amount of DNA
 Therefore the cell must synthesize more histones to
accommodate this increase

 Synthesis of histones occurs during the S phase

 Histones assemble into octamer structures
 They associate with the newly made DNA very near the replication fork

 Thus following DNA replication, each daughter strand

has a mixture of “old” and “new” histones
 Refer to Figure 11.22

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11-73
 Newly made
histone proteins

Figure 11.22
11-74
 Telomeres and DNA Replication
 Linear eukaryotic chromosomes have telomeres at both
ends

 The term telomere refers to the complex of telomeric

DNA sequences and bound proteins

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11-75
 Telomeric sequences consist of
 Moderately repetitive tandem arrays
 3’ overhang that is 12-16 nucleotides long

Figure 11.23

 Telomeric sequences typically consist of

 Several guanine nucleotides
 Often many thymine nucleotides
 Refer to Table 11.5
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11-76
11-77
 DNA polymerases possess two unusual features
 1. They synthesize DNA only in the 5’ to 3’ direction
 2. They cannot initiate DNA synthesis
 These two features pose a problem at the 3’ end of
linear chromosomes

Figure 11.24

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11-78
 Therefore if this problem is not solved
 The linear chromosome becomes progressively shorter with
each round of DNA replication
 Indeed, the cell solves this problem by adding DNA
sequences to the ends of telomeres
 This requires a specialized mechanism catalyzed by the
enzyme telomerase
 Telomerase contains protein and RNA
 The RNA is complementary to the DNA sequence found in the
telomeric repeat
 This allows the telomerase to bind to the 3’ overhang

 The lengthening mechanism is outlined in Figure 11.25

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11-79
 Step 1 = Binding

The binding-
polymerization- Step 2 = Polymerization
translocation cycle can
occurs many times

This greatly lengthens

one of the strands
Step 3 = Translocation

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

RNA primer
Figure 11.25
11-80
 DNA Proof-reading and Repair
• 3 tahapan repair yang
umum terjadi:
excission,
polymerization, and
sealing
• Namun demikian ada
banyak sekali sistem
DNA repair tergantung
jenis kesalahannya (see
next slide)
Siklus Sel
 Phases of the Cell Cycle
• The cell cycle consists of
– Interphase – normal cell activity
– The mitotic phase – cell divsion
INTERPHASE

Growth
G1 (DNA synthesis)

Growth
G2
 Cell Division
• An integral part of the cell cycle
• Results in genetically identical daughter cells
• Cells duplicate their genetic material
– Before they divide, ensuring that each daughter cell
receives an exact copy of the genetic material, DNA
(DNA Replication)
 Eukaryotic Cell Cycle
The eukaryotic cell cycle has 5 main phases:
1. G1 (gap phase 1)
2. S (synthesis)
3. G2 (gap phase 2) interphase
4. M (mitosis)
5. C (cytokinesis)
The length of a complete cell cycle varies greatly
among cell types.

41
 Interphase
Interphase is composed of:

G1 (gap phase 1) – time of cell growth

S phase – synthesis of DNA (DNA replication)

- 2 sister chromatids are produced

G2 (gap phase 2) – chromosomes condense

42
 Interphase
Following S phase, the sister chromatids appear
to share a centromere.
In fact, the centromere has been replicated but
the 2 centromeres are held together by
cohesin proteins.
Proteins of the kinetochore are attached to the
centromere.
Microtubules attach to the kinetochore.

43
 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

Cohesin Chromatid
proteins
Centromere
region of
chromosome

Kinetochore
Kinetochore
microtubules
Metaphase
chromosome
44
 Interphase
During G2 the chromosomes undergo
condensation, becoming tightly coiled.

Centrioles (microtubule-organizing centers)

replicate and one centriole moves to each
pole.

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