Chapter 11: Problem 7
In Kornberg's initial experiments, it was rumored that he grew E. coli in Anheuser-Busch beer vats. (Kornberg was working at Washington University in St. Louis.) Why do you think this might have been helpful to the experiment?
Chapter 11: Problem 7
In Kornberg's initial experiments, it was rumored that he grew E. coli in Anheuser-Busch beer vats. (Kornberg was working at Washington University in St. Louis.) Why do you think this might have been helpful to the experiment?
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Get started for freeDescribe the role of \(^{15} \mathrm{N}\) in the Meselson-Stahl experiment.
Prokaryotic Okazaki fragments are in the range of 1200 nucleotides, while eukaryotic fragments are much shorter, more in the range of \(100-150\) nucleotides. Balakrishnan and Bambara (2013) suggest that the shorter length of Okazaki fragments is determined by nucleosome periodicity. Design an experiment to determine whether or not the length of Okazaki fragments in eukaryotes is dependent on nucleosomes being present on \(\mathrm{J}\)
DNA polymerases in all organisms add only \(5^{\prime}\) nucleotides to the \(3^{\prime}\) end of a growing DNA strand, never to the \(5^{\prime}\) end. One possible reason for this is the fact that most DNA polymerases have a proofreading function that would not be energetically possible if DNA synthesis occurred in the \(3^{\prime}\) to \(5^{\prime}\) direction. (a) Sketch the reaction that DNA polymerase would have to catalyze if DNA synthesis occurred in the \(3^{\prime}\) to \(5^{\prime}\) direction. (b) Consider the information in your sketch and speculate as to why proofreading would be problematic.
You have generated a mutant strain of eukaryotic cells that constitutively express proteins required for translesion DNA synthesis (TLS). Would these cells have a mutator phenotype? Explain. One of the strains that you are working with shows an additional mutation whereby the processivity of a TLS polymerase is increased. What would be the consequence of this mutation?
In this chapter, we focused on how DNA is replicated and synthesized. We also discussed recombination at the DNA level and the phenomenon of gene conversion. Along the way, we encountered many opportunities to consider how this information was acquired. On the basis of these discussions, what answers would you propose to the following fundamental questions? (a) What is the experimental basis for concluding that DNA replicates semiconservatively in both prokaryotes and eukaryotes? (b) How was it demonstrated that DNA synthesis occurs under the direction of DNA polymerase III and not polymerase I? (c) How do we know that in vivo DNA synthesis occurs in the \(5^{\prime}\) to \(3^{\prime}\) direction? (d) How do we know that DNA synthesis is discontinuous on one of the two template strands? (e) What observations reveal that a "telomere problem" exists during eukaryotic DNA replication, and how did we learn of the solution to this problem?
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