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Rockefeller scientists first to reconstitute the DNA 'replication fork'
Date:7/9/2014

y the DNA sequence, but instead lies in modifications to proteins associated with the DNAis passed along to the daughter cells after DNA replication. Yet exactly how that occurs remains a mystery. Another unknown is what happens when the replication fork encounters an area of damaged DNA as it travels down the length of DNA.

"Diseases, such as cancer, often arise from DNA damage or defects in episomal inheritance, so these findings could have direct relevance to these fields," O'Donnell says. "There are plenty of hypotheses about the mechanics of DNA replication, but until now the process could not be studied using a defined system with pure proteins."

The replication fork is assembled as a complex of numerous proteins, one of which is an 11-subunit collective called CMG that unwinds and separates the DNA into two individual strands. The emerging replication fork looks much like a zipper opening, with CMG in the role of a zipper slider and the individual strand appearing like the two rows of teeth of the open zipper.

Each of these strands then becomes the templates for daughter copies. The act of synthesizing a new complementary strand to match the templates is performed by two different polymerase enzymes, which match each complementary subunit of DNA (the nucleotide "letters" that make up the genetic code) to the chain, resulting in a new double-stranded DNA molecule. These enzymes are known as polymerase epsilon (Pol epsilon) and polymerase delta (Pol delta), and the O'Donnell laboratory sought to examine how they attach to DNA to perform their task.

One of the chief features of the replications fork is its essential asymmetry. Because the two strands of double-stranded DNA are complementary, they fit together head to tail (in biochemical terms, the 5' end to the 3' end), so that the head of one strand is attached to the tail of the other. New DNA can only be synthesized in one direction (5' to 3'). This leads to a traffic pro
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Contact: Franklin Hoke
fhoke@rockefeller.edu
212-327-8998
Rockefeller University
Source:Eurekalert

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