Navigation Links
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 enzyme can reel in the DNA and in the process kick off recombination proteins," said Ha, who also is a Howard Hughes Medical Institute investigator.

When PcrA reaches the end of its DNA rope, it releases it and starts the reeling in process over again, removing any additional problematic proteins that have bound to the damaged DNA as it reels.

By using FRET, a technique Ha developed, the team also was able to answer another question about PcrA: How consistent is its motor function? Researchers agree that on average, PcrA moves one DNA unit, called a base pair, for each unit of cellular energy it uses, called ATP. But because researchers traditionally study the enzyme in relatively large samples, broad distributions of data have led to conflicting views on whether the helicase moves in uniform steps or those of varying lengths even up to six base pairs per ATP.

Since FRET is a single-molecule technique, the researchers were able to document a single enzyme's function, step by step, and found that PcrA does, in fact, move in uniform steps of one base pair per ATP.

Next, the team plans to create a reaction environment more similar to that in vivo, using three and four colors of FRET dyes to measure activities of multiple proteins simultaneously. They are also working toward understanding why helicase moves only in one direction.

"This is an ideal marriage of a new technology and an interesting biological problem," Ha said.


'/>"/>

Contact: Liz Ahlberg
eahlberg@illinois.edu
217-244-1073
University of Illinois at Urbana-Champaign
Source:Eurekalert  

Related biology news :

1. NC State researchers get to root of parasite genome
2. Researchers find animal with ability to survive climate change
3. Researchers find an essential gene for forming ears of corn
4. Researchers note differences between people and animals on calorie restriction
5. Researchers study acoustic communication in deep-sea fish
6. Researchers discover that growing up too fast may mean dying young in honey bees
7. Researchers study how pistachios may improve heart health
8. UI researchers find potentially toxic substance present in Chicago air
9. Researchers develop new self-training gene prediction program for fungi
10. Case Western Reserve University researchers track Chernobyl fallout
11. Childrens National researchers develop novel anti-tumor vaccine
Post Your Comments:
*Name:
*Comment:
*Email:
Related Image:
Researchers advance understanding of enzyme that regulates DNA
(Date:1/25/2016)...  Glencoe Software, the world-leading supplier of image data ... provide the data management solution OMERO Plus for the ... Photo - http://photos.prnewswire.com/prnh/20160125/325328LOGO ... Phenotypic analysis measures the characteristics and behavior of ... states such as health and disease, the presence or ...
(Date:1/22/2016)... 2016 http://www.researchandmarkets.com/research/p74whf/global_biometrics ... "Global Biometrics Market in Retail Sector ... --> http://www.researchandmarkets.com/research/p74whf/global_biometrics ) has announced ... Market in Retail Sector 2016-2020" report ... Research and Markets ( http://www.researchandmarkets.com/research/p74whf/global_biometrics ) has ...
(Date:1/20/2016)... A market that just keeps on growing. Molecular Diagnostics ... genomics knowledge. Learn all about it in this new ... trends are pushing market growth and company valuations. Trends ... pathogen evolution - next generation sequencing - emergence of ... the role of genetic material in Disease and Health ...
Breaking Biology News(10 mins):
(Date:2/11/2016)... and GERMANTOWN, Maryland , ... Frankfurt Prime Standard: QIA) today announced the introduction of ... gene expression profiling, expanding QIAGEN,s portfolio of Sample to ... researchers to select from over 20,000 human genes and ... between genes, cellular phenotypes and disease processes. --> ...
(Date:2/11/2016)... Feb. 11, 2016  Dovetail Genomics™ LLC today announced ... program for a planned metagenomic genome assembly service. ... metagenomic genome assembly method in a talk on Friday, ... & Technology conference in Orlando, Fla. ... complex datasets is difficult. Using its proprietary ...
(Date:2/11/2016)... , ... February 11, 2016 , ... ... its new stem cell treatment clinic in Quito, Ecuador. The new facility will ... trauma applications to patients from around the world. , The new GSCG ...
(Date:2/10/2016)... Feb.10, 2016 ASAE is introducing a hybrid ... Companies (AMC) the option of joining or renewing through ... determined by staff size, every employee in any size ... and reap all available member benefits.   ... organizational membership options will allow organizations of any size ...
Breaking Biology Technology: