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5 3
3 5
3-CCTACGCA-5
Two ssDNA molecules joined by
standard base-pairing rules
In dsDNA, the strands are always
complementary.
TB
Bacterial DNA replication
DNA synthesis using a DNA template
Complementary base pairing
(A=T, GC) determines the sequence
of the newly synthesized strand.
TB
bacterial DNA replication
origin (start point) bidirectional
bacterial
chromosome
TB
two replication forks
theta
structure
TB
semi-conservative
*
*
*
+
* TB
Bacterial DNA replication
Key Enzymes
helicase
ssDNA binding protein
primase
DNA polymerase III
DNA polymerase I
DNA ligase
TB
helicase
Unwinds duplex DNA
TB
SUPERKOIL-TOPOISOMERASE
ssDNA binding protein
binds to and stabilizes ssDNA
TB
Template
A sequence of DNA or RNA that directs
the synthesis of a complementary
sequence
Primer
The initial segment of a polymer that is to
be extended on which elongation depends
primase
synthesizes a short RNA primer
using a DNA template
primase
RNA primer
(a short starting sequence
made of RNA) TB
Polimerase
Memerlukan primer dan cetakan DNA
Polimerisasi diperpanjang pada 3
Aktivitas eksonuklease 3 -5, berfungsi
sebagai proofreading
Aktivitas eksonuklease 5-3 untuk
menghilangkan primer
DNA polymerase III
Synthesizes DNA from a DNA
template and proofreads
TB
DNA polymerase I
TB
DNA ligase
Joins DNA strands together by
forming phosphodiester bonds
DNA ligase
TB
replication fork
5'
lagging strand
3'
5'
leading strand
template strands
3' TB
Leading strand 5'
synthesis
RNA primer
helicase
ssDNA binding proteins
3' TB
5'
DNA polymerase
helicase
ssDNA binding proteins
3' TB
Leading strand synthesis 5'
helicase
DNA
ssDNA binding proteins 3' TB
Lagging strand synthesis
(discontinuous) Okazaki
fragment
3' (~1000 bases)
pol III
(primase)
5'
helicase
ssDNA binding proteins
3' TB
Primer removal
3' pol III
5'
pol I
5 to 3
pol I exonuclease
activity
TB
Proofreading
Pol III removes misincorporated bases
using 3' to 5' exonuclease activity
This decreases the error rate to about
10 per base pair inserted
-10
TB
Ligase DNA
Menyambung dua fragmen Okasaki
dengan membentuk ikatan fosfodiester
antara 3-OH fragmen 1 dengan 5-P
fragmen 2
Ligation
DNA ligase
TB
KEPERLUAN REPLIKASI DNA
TEMPLATE (CETAKAN)
PRIMER : 3-OH - PERPANJANGAN
PREKURSOR : dNTP
Enzim : polimerase DNA, helikase,
primase, SSBP, ligase
5
3
5
Lokus awal replikasi (Ori)
INISIASI
oxyS/fhlA in E. coli
oxyS RNA transcript
induced by stress
fhlA transcriptional
activator site
oxyS/fhlA complex
binds via two loop-loop
interactions
Bacterial chromosomes
Plasmid antisense RNAs are generally cis-
encoded
Implies complete Watson-Crick
complementarity
Bacterial chromosomes contain trans-
encoded antisense RNAs
Not necessarily complete complementarity
Often stress-related control systems
Prokaryotic vs. Eukaryotic
5' 3'
template TB
Telomeres
repetitive DNA at the end of linear
eukaryotic chromosomes
Example
(GGGGTT)n n = 20 - 200
Telomerases are
composed of protein and
an RNA molecule that AACCCCAAC
AACCCCAAC
GGGGTTGGGGTT
5'
telomerase
AACCCCAAC
GGGGTT GGGGTT GGGGTT
5'
primase
5'
pol I
ligase
telomeric repeats
Telomerase Demo created by Dr. Donald F. Slish at SUNY, Plattsburgh.
Note -on the telomerase enzyme, T's are used in the RNA template. This was an inadvertent
error. Since this template is RNA, these should be U's
3 5
5 3
3 5
3 5
3 5
5 3
3 5
5 3
3 5
First cycle
5 3
3 5
5 3
3 5
First cycle
5 3
3 5
5 3
3 5
Second Cycle
Second Cycle
Second Cycle
Third cycle
Third cycle
Third cycle
From 3rd cycle onwards this species will predominate