DNA Replication DNA synthesis which occurs within the nucleus

Enzyme: DNA-directed DNA polymerase

3 Models of DNA Replication:
1. Conservative Replication the parent DNA remains intact while the both strands
of the second DNA are newly synthesized.
2. Semi-conservative Replication Each strand of both daughter DNA molecules
came from the parent DNA.
3. Dispersive Replication Each strand of both daughter DNA molecules contains a
mixture of old and newly synthesized parts.
Features of DNA replication
1. Semi-conservative
2. Ordered and sequential
3. Uses activated substrates
4. Semi-discontinuous
5. Accurate
Three Steps for Replication
1. Initiation
a. Recognition of origin of replication by specific proteins
b. Formation of replication fork

Proteins involved in the formation of replication fork

Dna A for identification of origin of replication
DNA Helicase unwinds the DNA
Single strand DNA binding protein prevents the reannealing of DNA
Topoisomerase relieves the torsional stress brought by the unwinding of the
DNA

2. Elongation
2 strands leading strand (3 5) and the lagging strand (5 3)
Synthesis is catalyzed by DNA polymerase III - synthesis new DNA strand in 5
to 3 direction
Requirements of DNA pol III to synthesized a DNA strand using DNA as a
template:
a. Polymerization requires activated substances (dTTP, dATP, dGTP, dCTP)
and Mg+2
b. Requires a primer because it can only add deoxynucleotides at the 3 OH
group of previous nucleotide
Primase a DNA-directed RNA polymerase that synthesized short RNA chains
which will be use as a primer
RNA primer + protein molecules at the replication fork = Primosome

Leading strand requires only one RNA primer

Lagging strand requires several RNA primers
New DNA strand on the lagging strand are synthesized on a semidiscontinuous fashion.
Synthesis of Okazaki fragments (small DNA fragments) on the lagging strand

3. Termination

DNA polymerase I removed the RNA primers (exonuclease activity: 3 5

and 5 3) and replaced with deoxynucleotide.
DNA Ligase Nick Ligation seals the nick gaps by forming the
phosphodiester linkage.