Werner's syndrome (WS), a rare autosomal recessive disease due to mutations in a WRN helicase gene, is caused by genome instability. Patients affected with WS exhibit predisposition to cancer, and premature aging, accompanied by development of bilateral cataracts, diabetes mellitus, and osteoporosis. WRN helicase is a critical factor in DNA repair by homologous recombination, which repairs damage due to double strand breaks ((DSB) and stalled replication forks. The goal of this project is to understand how WRN functions in this pathway by examining the response of WRN to DNA damage; and its interactions with other factors. In normal cells and cells responding to DNA damage, WRN nuclear foci are clearly visible by immunofluorescent microscopy. It is not known whether this localization of WRN depends upon helicase activity, interactions with other factors, or both.
The aims of this proposal are to determine whether WRN localization is dependent on helicase activity and/or interaction with RAD52, an important repair factor. I will use fluorescent microscopy to analyze localization of wild-type WRN, mutant WRN which lacks helicase activity, and mutant WRN which is unable to interact with RAD52; and compare localization in cells growing under normal conditions and cells treated with agents that cause DNA damage. These experiments will provide us with a clear picture of how WRN localizes in a living cell and if localization is dependent upon intrinsic enzymatic activity or interactions with other proteins. ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Predoctoral Individual National Research Service Award (F31)
Project #
5F31GM073318-03
Application #
7116728
Study Section
Minority Programs Review Committee (MPRC)
Program Officer
Gaillard, Shawn R
Project Start
2004-09-16
Project End
2009-09-15
Budget Start
2006-09-16
Budget End
2007-09-15
Support Year
3
Fiscal Year
2006
Total Cost
$31,846
Indirect Cost
Name
University of Washington
Department
Biochemistry
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195