Chapter 12: DNA Replication

Problems:

COMPREHENSION QUESTIONS

1. What is semiconservative replication?

2. How did Meselson and Stahl demonstrate that replication in E. coli takes place in a semiconservative
manner?

3. Contrast DNA synthesis on the leading and lagging strands of DNA. Why is the mechanism different on the
two strands?

4. List the different proteins and enzymes taking part in bacterial replication. Give the function of each in the
replication process.

5. Use the key below to answer questions a-d.

a. DNA polymerase III

b. initiator proteins
c. helicase
d. primase
e. DNA ligase

a. During DNA replication, newly synthesized DNA contains many mistakes (mismatched base pairs). This
suggests a mutation in the gene coding for ___?
b. During DNA replication, the RNA primers between Okazaki fragments are removed but the DNA fragments
are never joined together. This suggests a mutation in the gene coding for ___?
c. During DNA replication, a replication bubble forms but synthesis of new DNA is never initiated. This
suggests a mutation in the gene coding for ___?
d. A prokaryotic cell is apparently unable to initiate replication (i.e. a replication bubble never forms). This
suggests a mutation in the gene coding for ___?

6. What similarities and differences exist in the enzymatic activities of DNA polymerases I and III? What is the
function of each DNA polymerase in bacterial cells?

7. In what ways is eukaryotic replication similar to bacterial replication, and in what ways is it different?

8. What is the end-of-chromosome problem for replication? Why, in the absence of telomerase, do the ends of
chromosomes get progressively shorter each time the DNA is replicated?

APPLICATION QUESTIONS

9. Suppose a future scientist explores a distant planet and discovers a novel form of double-stranded nucleic
acid. When this nucleic acid is exposed to DNA polymerases from E. coli, replication takes place continuously
on both strands. What conclusion can you make about the structure of this novel nucleic acid?

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10. A circular molecule of DNA contains 1 million base pairs. If the rate of DNA synthesis at a replication fork
occurs at a rate of 100,000 nucleotides per minute, how much time will theta replication require to completely
replicate the molecule, assuming that theta replication is bidirectional? How long will replication of this circular
chromosome take by rolling-circle replication?

11. A bacterium synthesizes DNA at each replication fork at a rate of 1000 nucleotides per second. If this
bacterium completely replicates its circular chromosome by theta replication in 30 minutes, how many base
pairs of DNA will its chromosome contain?

12. What would be the effect on DNA replication of mutations that destroyed each of the following activities in
DNA polymerase I?

a. 3’ to 5’ exonuclease activity
b. 5’ to 3’ exonuclease activity
c. 5’ to 3’ polymerase activity

13. How would DNA replication be affected in a bacterial cell that is lacking DNA gyrase?

14. If the gene for primase were mutated so that no functional primase was produced, what would be effect on
theta replication?

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