The lack of functional expression of proteins at the cell surface due to misfolding has been linked to number of genetic diseases, including familial hypercholesterolemia and cystic fibrosis. However, the molecular mechanisms responsible for correct folding of membrane proteins in the ER remain poorly understood. The goal of the proposed studies is to investigate the molecular mechanisms of folding and surface expression of CFTR, a polytopic integral membrane protein. Experiments outlined in the first aim will focus on identification of proteins involved in CFTR folding/surface expression. For this purpose a genetic screen will be carried out in mammalian cells to identify high copy extra genic suppressors of deltaF508 CFTR processing defect. Isolation of CFTR folding modulators and characterization of their interactions with both the wild type and mutant CFTR should provide insights into the cellular regulation of CFTR surface expression.
The second aim i s to identify second-site mutations within deltaF508 CFTR molecule that can restore its proper folding and surface expression. These experiments will utilize PCR-based mutagenesis combined with expression cloning in mammalian cells. Understanding of membrane protein folding mechanisms is central for issues of cell biology and new insights into the regulation of CFTR folding and surface expression can be instrumental in the development of treatments for genetic diseases caused by protein folding/trafficking defects.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32DK009833-03
Application #
6176449
Study Section
Biological Sciences 2 (BIOL)
Program Officer
Hyde, James F
Project Start
2000-08-01
Project End
Budget Start
2000-08-01
Budget End
2001-07-31
Support Year
3
Fiscal Year
2000
Total Cost
$39,232
Indirect Cost
Name
Stanford University
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305