DNA Replication

DNA is semiconservative
o Semiconservative replication: A mechanism of DNA replication in
which each of the two strands of parent DNA is incorporated into a
new double stranded DNA molecule.
o Involves separating the two parent strand and building a new
complementary replacement strand for each

DNA Replication: The process

Step 1: Strand Separation
DNA must first unwound each other (replication origin is the starting point)
o Replication Origin: A specific sequence of DNA that acts as a starting
point for replication
Helicase binds to the starting point and starts to unwind the two DNA
strands by breaking the hydrogen bonds between the complementary base
pairs.
o Helicase: A replication enzyme that separates and unwinds the DNA
strands
As strands start to separate, they for a y shape known as the replication fork.
o Replication Fork: The point of separation of the two parent DNA
strands during replication
An enzyme called Topoisomerases prevents DNA strands from being twisted
and tangled.
o It relieves tension by cutting one or two of the strand near the
replication fork
o Topoisomerase: An enzyme that relieve tension caused by the
unwinding of parent DNA; they cleave one or two of the DNA strand,
allow the strands to untwist and then rejoin the strands
DNA has the tendency to separate and bind together again so to prevent that
SSBs prevents the H bonds from reforming and annealing.
o SSBs (single-strand binding protein): A replication enzyme that
prevents parent DNA strand from annealing to each other once they
have been separated by helicase

Step 2: Building Complementary Strands
New nucleotides are joined by a group of enzymes called DNA polymerase.
Both prokaryotes and eukaryotes have DNA polymerase
Prokaryotic cell add DNA from 3 to 5 end while eukaryotic cell add DNA
from 5 to 3 end
Builds new strand by using nucleoside triphosphates
o a building block and energy source for replicating DNA
RNA primase begins the replication stage and set small complementary RNA
segment at the beginning of the replication fork
o RNA Primase: A replication enzyme that produces RNA primers
About 10-60 RNA are produced also known as RNA primer
o RNA Primer: A replication molecule that acts as a starting point for
replication
Once RNA primer is set, DNA polymerase III starts adding DNA nucleotides to
the RNA primer
o DNA polymerase III: A prokaryotic replication enzyme that builds
new DNA strands from nucleotides
o Can only build from 5 to 3 end
Strands start building in opposite direction
The DNA polymerase III that is building towards the replication for is able to
continuously build (no RNA Primer needs to keep adding to RNA primase)
o Known as the Leading strand
o Leading Strand: The DNA strand that is copied in the direction
towards the replication fork
On the other strands, the DNA polymerase III is replicating away from the
replicating fork.
o Lagging Strand: The DNA strand that is copied in the direction away
from the replication fork
RNA Primase will attach itself to part of the lagging strand and will be
replicating in pieces called Okazaki fragments
o Okazaki fragments: The piece of new DNA on the lagging strand
Lagging strand built in pieces and is not a continual process since it is made
in pieces
Each fragment extend in the 5 to 3 direction, it eventually runs into the RNA
primer attached to the Okazaki fragment ahead
DNA polymerase I removes the RNA primer/nucleotides one at a time and
replaces it with DNA nucleotides
o DNA Polymerase I: A prokaryotic replication enzyme that fills in
gaps in the lagging strand between Okazaki fragments; also
proofreads the final strand
DNA ligase connects the new DNA nucleotides and Okazaki fragments
together
o DNA Ligase: An enzyme that catalyzes the formation of a
phosphodiester bond between two DNA strands, as well as between
Okazaki fragment.

Step 3: Dealing with Errors during DNA Replication
A DNA polymerase proof reads the strand and corrects the errors (usually
base pair match)
DNA Polymerase III cannot move on if there is a error in the strand
Composed of proteins and enzymes including DNA polymerase I and
polymerase II
DNA polymerase II is a slow enzyme that works to repair damages to DNA
o DNA polymerase II : A prokaryotic replication enzyme that repairs
damage to DNA, including damage that occurs between replication
event
Mismatch base pairs will not be able to bond correctly and will distort the
shape of the DNA