Side note:

Model organism: Easy and cheap to keep, short generation time, habloid (recessive gene mutation), can make tetratypes
(crossmutations), can keep plasmids

DNA replication

Watson & Crick 1953 in Nature: "A Structure for

Deoxyribose Nucleic Acid“:

“It has not escaped our notice that the specific pairing
we have postulated immediately suggests a possible
copying mechanism for the genetic material”.
Conservative, semi-conservative or dispersive DNA replication?
Meselsohn & Stahl experiment
Nitrogen is found in the bases of cells, proteins (the backbone) and more
 Meselsohn og Stahl

15N

14N
Conservative?

Semi-conservative?

Dispersive?
15N – shift to 14N
Conclusion of the Meselsohn Stahl experiment:

DNA replication is “semi-conservative”

DNA is kept together by hydrogen bonds between purines and pyrimidines and stacking forces generating the double helix shape
 DNA synthesis is directed in the 5' to 3'

pyrophosphate = ppi

By Michał Sobkowski - Own work, CC BY 3.0,

https://commons.wikimedia.org/w/index.php?curid=14825501
DNA replication in direction of 5’ towards 3’
DNA polymerase

Moving from 3' to 5' and synthesise from 5' to 3'

A fork replicates both strands simultaneously

paradox = antiparallel
 Replication in both 5’-3’ and 3’-5’ directions?????

There is no DNA polymerase able to synthesize in the 3’-5’ direction

DNA synthesis is discontinuous on
the lagging strand
And continuous on the leading strand

The short “Okazaki”-fragments on

lagging strand are ligated together by
DNA ligase.
DNA synthesis is discontinuous on the lagging strand
Leading strand is continuous
DNA ligase covalently links the pieces
of the lagging strand pieces
DNA polymerase is dependent on a perfect
basepair for synthesis
= proof reading activity

Hence DNA polymerase cannot initiate

synthesis of DNA without a primer = has to read something before it can start

DNA synthesis is initiated by RNA synthesis

DNA polymerase initiates from short RNA-primers

3' 5'

5' 3'
Two forks move in opposite direction from the place of initiation
DNA helicase exposes single stranded DNA
Use ATP to unwind double stranded DNA
 Replication protein A

SSB/RPA proteins bind to single stranded DNA

prevent refolding or misfolding of single stranded DNA
Synthesis of the lagging strand
 A replication fork
Work in mitosis and meiosis

RPA
Proteins involved with DNA synthesis:
DNA helicase
DNA polymerase
Primase = put down RNA primer
PCNA
DNA ligase
Topoisomerase = reduce torsion tension/stress by cutting the DNA single stranded or double stranded

And ?

endonuclease = release RNA bits of lagging strand and polymerase can replace with DNA
Specialized enzymes for initiation of replication = the origin of replication

RNR = ribonucleotide redoxtase

= reduce the phosphate groups in the ribonucletide -> Make the building blocks for the DNA synthesis
A replicated condensed chromosome

How many DNA strands does it contain?