Replication

Density -

transfer experiments !
" "
DNA
Type semi conservative

I 4N
'
I Stahl centrifugation
'

Experiment 1 : Meselson 5N

"
semi + semi = semi + N

" "

Experiments :
Taylor ; radioactive 311 -

thymidine → Black spot

: colchicine inhibit Anaphase separation

ñÑw semi ñÑiwwÑoD Black spot ÑnwwtaiÑ Black spot
)
*

of replication E. coli
origin
origins of replication Corio )
nucleotide
Eukaryote Prokaryote 245
-

•
multiple origin •
single origin
- 82 qnÑo 13 -

bp I g- bp

Because
~

l / \
aroniiqnioieuiiaiwo

seqbasepai
DNA

initia+ionpr◦+
L
Replication oh
13 -

bp g- bp

ÑA=T 6h08AM sequence a

times
manor breakdown ironic #

binding site
replication bubble
To qdi%me9Ñ
"

Enzyme
"

ÑAmsms Replication

2
I
 Replication process
Initiation of Rep .

1. start Oric .

2. Bit !aA protein a molecules

binding 9- bp
Mi 20-40 polypeptides in Orio

rhinitis 13 bp frissons an
-

2nd !
3. Dna B ( DNA helicase) & Dirac protein
bÑ1anÑqaÑwÑww-Ñw Replications ble
↳ a bidirectional
replication forks

Role of Topo im erase in DNA Rep .

•
dna B ( DNA helicase) → Unwind of DNA

↳ using AIEnergY
• SSB →
ÑosÑwmÑU§nÑw v00 DNA
I

↳ ssBñÑÑ stimulates miioorihsvoo monomers

no
binding 1st

• DNA
topoisomerase → Protect minimising DNA ;TnwmiuqwiÑw9ÑÑownñornÉwo
(
DNA
topoisomerase I l
DNAtopoimerase I
•
LE Energy ! ◦
% Energy ! [ ATP )
◦ oin DNA intowww.UN.wniwioioiofwiuafowioiw ◦ oinks 268 www.otniuriioohsetots :

Template strand
j
'

Re formation vos bond

phosphodiester
◦ - g-

{Ubbuuvos
DNA helix & DNA topoisomerase of g-

Extension of polynucleotides chain

Initiation of DNA chain with RNA
primer
◦ DNA polymerases win nucleotide
◦
DNA primase Athos RNA primer Murthy
2
◦
Nucleophilic Ñw 3- OH Unico RNA primer P

bÑofñÑ ( 5- OH )9Ñ
polymerase Norio ◦

synthesis b- → 3 Kimani ! ) .

Eukaryote
/ prokaryote "

'
'

④ -5
'
Ñ 09607 MM @ OH -3

pm 10-60 nucleotides • to nucleotide only ! Mosan ④ -5 'd

Energy ÷
→
É⑤-① 1
OH
'
3

↑
¥⑧ ⑧
 Activity of DNA polymerase
◦
polymerase activity ◦
Exonuclease activity
long Short crinoid
Wonga)
-
-

using Energy
-

not
⇐ using Energy
ÑnÑoW0nqn
'
-
→ 3 -
OH or 5- P

DNA
polymerase III of E. coli

catalytic 3 core → ✗
,
E
,
O

'Ñwg
'

lock
p subunit of score
-
o
r

[Fidelity ÑDÑATepiiaTeÑ
◦

; aoiwiiaiwiims Replicated DNA

replication fork
o
◦ - o - o -
- o

◦ error rate of 10 -5
to 10-4 → error I / 104 Base leading stand } '

MMÉ
continuous
"
o
"

proofreading mechanism
"

→
protect Error ! luo% I / 10° Base % ↳ Discontinue
→

lagging stand
•
Exo nucleus
activity → Fidelity of DNA Replicated
I
%
•

leading stands Lagging stands

Topoisomerase
•
Deny RNA primers
• →
protect DNA
winding "

DNA Inuit raw DNA

poly #
Unga replicated
→ Fork →

protect DNA
SSB
winding DNA
→
e
ligase
◦

→ bmiÑDNAdWbÑod % ATP
→
Energy an NAD or

lagging strand

•
Exonuclease ; 5 → 3 • DNA
ligase Wario Okazaki
fragment
food
phosphodiester bond
→ DNA poly I 607 RNA primer ooh

◦
Polymerase ; b-→ 3

"

→ Enzyme a. 607 Okazaki fragment 2h6m WÑ

( 3- OH)
 Repli some
*
Lagging strand ; DNA 0 :ÑmivabÑᵈoÑÑnñ DNA poly II
contorts RNA primer
'

Mantovani structure Ñ DNA wenviw move

loop lÑonnwiwmr Replicated

termination

•
Ya termination of Replicated wow
ga block Mrs :u= cqnr.mn )
•
Topoim erases →
Negative supercoiling

Rolling -

circle of replication

¥s ↳ Stanger replication plasmid

in

Virus / Bacteria unwon

◦
Eukaryotic chromosome replicated
multiple origin
} oianrnconnectriwiotunivyifn
→

→
multiple terminal no

DNA polymerase
prokaryote I

Polymerase II

compare Eukaryote Prokaryote

Histone
•
assembly
Clock DNA)
DNA PNCA
Sliding Clamp Eukaryote .

43 subunit ) initiation replicated

-

•
polymerase ✗ -

Ribonuclease
Exo nucleus I
a
HI DNA
MY •
Polymerase 8 -

complete replicated lagging strand

( RNA primer → Dna,

FEN -7
•
Polymerase E -

carry
out polymerisation lead
ding strand
Nucleosome =
histon Ww ÑU DNA

Nap -1 histone
binding CAF -1
•
m an

bÑÑÑ histone oiondu DNA

'

telomerase Telomere

◦ mint was today 680 Replicated

◦ Telomerase → Enzyme ÑoÑÑVmumowoÉw
into replicated

→
Hybrid ; DNA tRNA

doing RNA primer winds in

gap
↳ oohs multiple repeat of telomere

◦ nuts Canete call ,

stem cell

telomere length and

aging
↳ nuts cell
ñiaiouj moi
↳ wound is somatic ( non telomerase)
-
→ iwaki : mon !
-

↳ stem cell