Recitation 11: DNA Replication
Learning Goals
Today’s Recitation:
(1) Explain DNA replication
(2) Describe the function of DNA polymerase, DNA helicase, primase, DNA ligase, and
topoisomerases during replication.
Big Picture: Understand that the Central Dogma describes information transfer in the cell
and includes the following three processes: DNA replication, transcription, and
translation.
DNA Replication
DNA replication occurs during the S phase (S = synthesis) of cell division when two
double-stranded DNA molecules are made from one double-stranded DNA (dsDNA)
molecule. Each daughter cell then receives one copy of newly synthesized dsDNA, which
is identical to that in the parental cell. DNA replication is semi-conservative and the newly
synthesized strand is antiparallel and complementary to the template strand. In the cell,
replication is continuous for one strand (leading strand) and discontinuous for the other
strand (lagging strand).
Test Yourself 1: In your own words, explain what semi-conservative replication means.
DNA Replication Enzymes
Replication starts at the origin of replication (ori) site. The DNA polymerase enzyme
catalyzes the addition of complementary bases to the 3’ OH end of the newly synthesized
strand. DNA polymerase adds complementary bases only to the 3’OH end of small RNA
primers that are made by the primase enzyme. These RNA primers are degraded after
replication. The gaps are sealed by adding the dNTPs via DNA polymerase, and the nicks
are sealed by DNA ligase.
During replication, the enzyme helicase unwinds the two strands of DNA such that they
can be accessed and replicated. Enzymes called topoisomerases remove the extra coils
that form as the new DNA is being synthesized and allow the DNA to unwind properly.
Problem 1: Replication Summary Complete the table below.
Answer
Subcellular organelle (s) in a eukaryotic cell where replication occurs is…
Monomer used to form DNA polymer?
Incoming monomers are added to…
What is the name of the bond that forms between two adjacent
monomers in a DNA strand?
Number of template strands needed to make a single DNA duplex?
Which direction (5’ → 3’ or 3’ → 5’) is the template DNA strand read?
Which direction (5’ → 3’ or 3’ → 5’) is the new DNA strand (both
leading and lagging) synthesized?
Problem 2: Replication Fork
Consider the DNA replication fork below.
A.) Would replication of the top strand be continuous or discontinuous?
B.) Draw and label the 5’ and 3’ ends of the daughter strand(s) made from the bottom strand.
C.) What is the function of a helicase? What direction would it be moving in this schematic?
Show with an arrow.
D.) If the DNA ligase protein is inactivated, which strand is going to be more affected?
Problem 3: Lagging Strand Synthesis.
The diagram below is a visual schematic (not real) showing how the leading and lagging
strands are synthesized during DNA replication.
Based on what you know about DNA replication, answer these questions:
a) In which direction (5' to 3' or 3' to 5') does DNA synthesis occur for both the leading and lagging
strands? How does this relate to the overall direction of the replication fork movement?
b) The DNA polymerase for the lagging strand synthesizes DNA faster than the leading strand
polymerase, however replication occurs at the same pace for both strands. Why does the lagging
strand polymerase have to be faster to maintain the same pace? In your explanation, name the
enzymes involved in synthesis of the lagging strand.
Fidelity of DNA Replication
Very few errors are introduced during DNA replication: only about one error in every 10 bases.
10
This low error rate is for the most part due to the ability of most DNA polymerases to proofread.
DNA polymerases can detect the wrong base and correct it during replication by either slowing
down and allowing an incorrect dNTP to dissociate from the active site, or by using a repair
exonuclease domain to cleave an incorrectly incorporated nucleotide from the new DNA strand.
Damaged bases can be removed from DNA by excision repair where the damaged base is flipped
out of the double helix and cleaved off by a repair enzyme. Mismatched base pairs can be repaired
by “mismatch repair” enzymes where the mismatched nucleotide (and some of its surrounding
nucleotides) are cleaved off so that the correct nucleotides can be added.
Problem 4: O6-Methyl-G
You are given the base pairing for A-T and C-G bases in the diagram:
O6-Methyl-G (O6MeG) is a guanine base that was damaged by a carcinogen that methylates DNA.
Compare C:G base pairing from the diagram given to the likely base pairing between a C and O6MeG.
Also predict the base pairing between O6MeG and T