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Researchers advance understanding of enzyme that regulates DNA
Date:8/20/2010

CHAMPAIGN, Ill. Thanks to a single-molecule imaging technique developed by a University of Illinois professor, researchers have revealed the mechanisms of an important DNA-regulating enzyme.

Helicase enzymes are best known for "unzipping" DNA for replication, but have many other functions for DNA repair and maintenance. The Illinois team focused on a particular bacterial helicase called PcrA involved in preventing unwanted recombination.

A DNA double helix consists of two strands twisted around each other. When one strand is damaged or breaks, the surrounding area is degraded, leaving a single-stranded region. Specialized proteins then start the process of recombination rebuilding the second strand using the intact DNA as a template.

"Recombination is essential for DNA repair, but if it runs amok, it causes problems," said U. of I. physics professor Taekjip Ha. "This helicase controls recombination by removing recombination proteins from the DNA."

Using a technique called single molecule fluorescence resonance energy transfer (FRET), Ha and his team were able to identify one of the mechanisms that PcrA uses to regulate recombination. The system uses two dyes that change in relative intensity depending on their proximities to one another. The researchers attached the two dyes to the opposite ends of the single-stranded DNA tail.

Helicases are motor proteins, a class of enzymes that use chemical energy to move along a DNA molecule like a train on a track. But using FRET, the researchers observed the two dyes gradually moving closer to each other, then flying apart, repeatedly. Instead of moving along the single-stranded tail, PcrA binds at the point of the break, where the double- and single-stranded regions meet. Then, it uses its motor function to "reel in" the tail, like a fisherman pulling in a rope.

"By combining the structure-specific binding of the enzyme to the DNA and the motor function, the e
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Contact: Liz Ahlberg
eahlberg@illinois.edu
217-244-1073
University of Illinois at Urbana-Champaign
Source:Eurekalert  

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