DNA

Cj Gutierrez

A. 3 Phases of Replication
1. Initiation
a) Recognition of Origin of Replication
b) Unwinding of double stranded helix
Helicase
o Enzyme that helps unwind that helix
DNA Gyrase
o A kind of topoisomerase
o Relieves torsional strain
SSB
o Stabilize unwound parental strand
c) Introduction of RNA primer
The primer is an oligonucleotide
It is only introduced at the start and provides a free 3 Hydroxyl which
reacts to the Phosphate group of the incoming nucleotide resulting in
the connection of the 2 nucleotides together
Found more in the Lagging Strand than in the Leading Strand
Primerase
o Enzyme that introduces the RNA primer
2. Elongation
o DNA Polymerase III
Uses dNTPs (deoxynucleoside triphosphate) and Mg 2+
o dATP + Mg2+
o dGTP + Mg2+
o dTTP+ Mg2+
o dCTP+ Mg2+
o This steps connects the partners (AT/AU and CG)
o Example
TEMPLATE GROUPING DNA STRAND
dTTP+ Mg2+
A
o Mg2+ removes/decreases the electronegativity of the O -
o This process uses a Triphosphate instead of a Monophosphate because the
reaction needs energy which is supplied by the Triphosphate or inorganic
phosphate
o Okazaki Fragment
RNA + DNA RNA + DNA = Found in Lagging Strand
Synthesis in lagging strand is discontinuous

3. Termination
Nicks Translation
o Removal of RNA primer
As RNA primer is removed, it is replaced by DNA
o Facilitated by the enzyme DNA polymerase I
o DNA ligase joins the Okazaki Fragment to the growing strand
 OTHER ENZYMES INVOLVED IN TRANSLATION

DNA polymerase III

o Main Enzyme
o Polymerization Rate: 250-1000 nucleotides/sec
o Processivity
DNA polymerase I
o Involved in Nicks Translation
o 5-3 Exonuclease
Needed to remove the RNA primer
o 3-5 Exonuclease
Proof reading to ensure 100% correct
Detects wrong nucleotides and cleaves it then replaces with correct
one

*Translation must have a high fidelity (Replication must be 100% correct)