5. A) DnaB is a helicase that loads as a ring onto one strand of the DNA and pus

Question

5. A) DnaB is a helicase that loads as a ring onto one strand of the DNA and pushes it apart from the second strand. This allows for 5’-3’ DNA unwinding and replication fork advancement that is necessary for initiation and elongation. If you create a temperature sensitive mutation in DnaB, what will be the effect on DNA replication (quick stop/slow stop mutation) at the non-permissive temperature? Why does this occur?

B) DnaA is a protein that binds to OriC (origin of replication) to initiate DNA replication. This protein must be made new with each round of replication. If you create a temperature sensitive mutation in DnaA, what will be the effect on DNA replication (quick stop/slow stop) at the non-permissive temperature? Why does this effect occur?

Explanation / Answer

effect of mutant Dna A & B on DNA replication

it has been experimentally studied that, the mutant Dna B protein showed mutation at one amino acid. this region is believed to be involved in interaction with Dna C. this mutation induces a temperature senitive conformational changes. These mutant have negligible primer RNA synthesis, ATPase activity and helicase activity at the non permissive temperature

Mutation in Dna A at non permissive temperature , showed that mutant show a fast - stop DNA synthesis phenotype typical of a replication fork block.

In contrast, ine mutant are able to complete at least oen round of DNA replication.

The effect of these mutation on the stability of replication complex was tested using a coprecipitation assay

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