DNA Repilication
DNA Repilication
Semi –conservative
Replication
Structure of DNA
SSBs
Keep separated DNA strands apart
by blocking hydrogen bonding
Keep the templates (single DNA
strands) straight
Replication Fork
The junction where the DNA strands
are still joined
2. Building complementary strands
Comp
THE PLAYERS
•DNA polymerase
•primase
•DNA ligase
DNA polymerase III
Takes free nucleotides found within
the cell and adds them in the 5’ to
3’ direction
The parent strand is used as a
template
Leading strand
The daughter strand that grows
continuously towards the replication
fork as the double helix unwinds
Occurs quickly
Requires a single RNA primer
Lagging strand
Built in short segments (in the 5’ to 3’
direction) away from the replication
fork
Requires many RNA primers
Lagging Strand Details
The 3’ to 5’ parent strand is a problem
for DNA polymerase since it must
synthesize in the 5’ to 3’ direction
Primase
Builds RNA primers (used to initiate DNA replication)
Okazaki Fragments
◻ Short fragments of DNA built by
DNA polymerase off of the RNA
primers
DNA polymerase I
Removes the RNA primers once they
have been used and replaces them with
the appropriate DNA sequence
DNA ligase
◻ Joins the Okazaki fragments into one
strand by the creation of
phosphodiester bonds
Link-leading vs lagging animation:
Leading Lagging
3. Linking Nitrogenous Bases
Nitrogen bases of nucleotides of
opposite strands (parent and daughter)
form new H bonds
Once bonds are formed, DNA twists
automatically into a double helix
QUALITY CONTROL
•
DNA polymerase acts as a proof-
reader by checking the newly
synthesized strand for any incorrectly
inserted bases
•If a mistake is found, act as an
exonuclease – & cut out the mistake
Summary
DNA helicase breaks H bonds and unwinds helix
Single-stranded binding proteins stabilize unwound DNA
DNA gyrase relieves tension
DNA is synthesized in a 5’ to 3’ direction by DNA
polymerase III from an RNA primer
DNA polymerase III builds in short segments called
Okazaki fragments
DNA polymerase I removes RNA primers and replaces
them with the appropriate nucleotide
DNA ligase joins fragments by completing the backbone
Animations
DNA Rep
DNA Rep2
Your Homework
Summarize Comparing DNA Replication in Euk and Prok. Know similarities
and differences
DNA Replication
Helicase
DNA gyrase
RNA primer
Ligase
DNA polymerase I
DNA polymerase II
DNA polymerase II
Primase
Correcting Errors during DNA
Replication
•
A human cell can copy all of its DNA in a couple of hours
with an error rate of 1 per billion nucleotide pairs.
•
The most common type of error is mispairing, such that a T
is paired with G. fig 5.20
•
Another error is caused by strand slippage during
replication. This can result in addition or omission of
nucleotides in the newly synthesized strand. Fig 5.21
•
Most polymerases have the ability to proofread. The
textbook stresses Pol II for some reason.
•
When an incorrect nucleotide is placed on the newly
synthesized strand, replication is stalled as the 3’ end is in
the wrong position for the next to attach.
•
The incorrect nucleotide is then excised (along with a few
correct nucleotides) and DNA replication can continue.
Proofreading Video
•
Mismatch repair is another mechanism that corrects errors
in both eukaryotes and prokaryotes that were not fixed
during proofreading. The mechanism can also detect
insertions and deletions that happens with strand slippage.
•
A protein complex recognizes and binds to the mismatched
base. A second complex cuts the DNA near the mismatch,
and more enzymes chop out the incorrect nucleotide and
adjacent patch of DNA.
•
A polymerase will then add the correct nucleotides.
Telomeres
•
Telomeres are long, non-coding, highly-repetitive
sequences of DNA at the ends of eukaryotic chromosomes.
•
They are necessary as DNA synthesis can only occur in the
5’-3’ direction.
•
After the final RNA primer is placed on the lagging strand,
the extension on the parent chain will not be matched with
nucleotides.
•
Thus each subsequent replication results in shortened
telomeres. If the telomeres where not present, then coding
regions (genes) would be shortened and most likely not
functional. This would lead to cell death.
•
Leading strand synthesis is normal
but look at the lagging strand.
•
Removal of the last RNA primer
results in a gap.
•
The length of the telomeres
is closely linked to the
lifespan of an organism.
•
Telomerase is enzyme that
can synthesize telomeric
regions that have been lost.
•
Telomerase activity high
during childhood when
tissues are growing rapidly.
Telomerase activity slows in
somatic cells (body) as
people age.
•
If we could maintain high telomerase activity we would
age much slower.
•
However, high telomerase activity is what allows for
cancerous tumors to grow. Cancer is unregulated cell
proliferation. High telomerase activity allows for the tumor
cells to continue to replicate as important genes would not
be lost.
Comparing DNA Replication
in Eukaryotes and Prokaryotes
•
Review this section on Pg 227 for your text.