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Ch. 11 - DNA Replication and Recombination
Chapter 11, Problem 20

Several temperature-sensitive mutant strains of E. coli display the following characteristics. Predict what enzyme or function is being affected by each mutation. Okazaki fragments accumulate, and DNA synthesis is never completed.

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Identify the process where Okazaki fragments are involved, which is DNA replication.
Recall that Okazaki fragments are short sequences of DNA nucleotides synthesized discontinuously and later linked together during DNA replication.
Understand that the enzyme responsible for linking Okazaki fragments is DNA ligase.
Consider that if Okazaki fragments accumulate and DNA synthesis is never completed, the enzyme likely affected is DNA ligase.
Conclude that the mutation affects the function of DNA ligase, preventing the joining of Okazaki fragments and completion of DNA synthesis.

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Key Concepts

Here are the essential concepts you must grasp in order to answer the question correctly.

Okazaki Fragments

Okazaki fragments are short sequences of DNA synthesized on the lagging strand during DNA replication. They are formed because DNA polymerase can only synthesize DNA in the 5' to 3' direction, necessitating the creation of these fragments as the replication fork opens. Understanding their formation is crucial for analyzing mutations that affect DNA synthesis.
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Steps to DNA Replication

DNA Polymerase

DNA polymerase is the enzyme responsible for synthesizing new DNA strands by adding nucleotides complementary to the template strand. In E. coli, DNA polymerase III is the primary enzyme for elongating the new DNA strand. Mutations affecting this enzyme can lead to incomplete DNA synthesis, as seen with the accumulation of Okazaki fragments.
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DNA Proofreading

Temperature-Sensitive Mutants

Temperature-sensitive mutants are organisms that exhibit normal function at one temperature but show a defect at a different temperature. In the context of E. coli, these mutations can affect enzymes like DNA polymerase, leading to issues in DNA replication at non-permissive temperatures. This concept is essential for predicting the effects of specific mutations on cellular functions.
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Mutations and Phenotypes
Related Practice
Textbook Question
Suppose that E. coli synthesizes DNA at a rate of 100,000 nucleotides per minute and takes 40 minutes to replicate its chromosome. (a) How many base pairs are present in the entire E. coli chromosome? (b) What is the physical length of the chromosome in its helical configuration—that is, what is the circumference of the chromosome if it were opened into a circle?
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Textbook Question

DNA supercoiling, which occurs when coiling tension is generated ahead of the replication fork, is relieved by DNA gyrase. Supercoiling may also be involved in transcription regulation. Researchers discovered that enhancers operating over a long distance (2500 bp) are dependent on DNA supercoiling, while enhancers operating over shorter distances (110 bp) are not so dependent [Liu et al. (2001). Proc. Natl. Acad. Sci. USA 98:14,883–14,888]. Using a diagram, suggest a way in which supercoiling may positively influence enhancer activity over long distances.

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Textbook Question
Several temperature-sensitive mutant strains of E. coli display the following characteristics. Predict what enzyme or function is being affected by each mutation. Newly synthesized DNA contains many mismatched base pairs.
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Textbook Question
Several temperature-sensitive mutant strains of E. coli display the following characteristics. Predict what enzyme or function is being affected by each mutation. No initiation occurs.
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Textbook Question
Several temperature-sensitive mutant strains of E. coli display the following characteristics. Predict what enzyme or function is being affected by each mutation. Synthesis is very slow.
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Textbook Question
Several temperature-sensitive mutant strains of E. coli display the following characteristics. Predict what enzyme or function is being affected by each mutation. Supercoiled strands remain after replication, which is never completed.
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