West Nile virus has become an increasing public health concern in the United States since its appearance in New York City in 1999. In order to gain a better understanding of the interaction of this virus with the infected host cell, we have performed gene microarray studies on WNV infected cells. The results of these studies suggested that the src-family kinase c-Yes may play a role in WNV infection. In agreement with this hypothesis, addition of the src-family kinase inhibitors PP2 and SU6656 to infected cells resulted in a 10-20 fold decrease in the amount of virus recovered in the culture supernatant 20 h after drug addition, and a rapid decrease of cell associated infectious virus soon (4 h) after drug addition. Interestingly, drug treatment does not result in a corresponding decrease in the amount of viral RNA within the infected cell, suggesting that c-Yes, or other src kinases, do not act on viral RNA replication, but at a later stage in the viral life cycle. Specific inhibition of c-Yes in an siRNA mediated """"""""knock-down"""""""" experiment also resulted in a decrease in recovered virus, indicating that c-Yes is indeed involved in WNV replication, although a role for other members of the src family cannot be excluded.
The aim of this application is to extend these studies to determine what members of the src kinases, other than c-Yes, are also involved in WNV replication, where in the viral life cycle the effect of c-Yes is exerted, and what substrates, viral or cellular, are phosphorylated by the kinase during infection. ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21AI061527-02
Application #
6915584
Study Section
Special Emphasis Panel (ZRG1-IDM-G (02))
Program Officer
Repik, Patricia M
Project Start
2004-07-01
Project End
2007-06-30
Budget Start
2005-07-01
Budget End
2007-06-30
Support Year
2
Fiscal Year
2005
Total Cost
$226,500
Indirect Cost
Name
Oregon Health and Science University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
096997515
City
Portland
State
OR
Country
United States
Zip Code
97239
Medigeshi, Guruprasad R; Hirsch, Alec J; Streblow, Daniel N et al. (2008) West Nile virus entry requires cholesterol-rich membrane microdomains and is independent of alphavbeta3 integrin. J Virol 82:5212-9
Medigeshi, Guruprasad R; Lancaster, Alissa M; Hirsch, Alec J et al. (2007) West Nile virus infection activates the unfolded protein response, leading to CHOP induction and apoptosis. J Virol 81:10849-60
Hirsch, Alec J; Medigeshi, Guruprasad R; Meyers, Heather L et al. (2005) The Src family kinase c-Yes is required for maturation of West Nile virus particles. J Virol 79:11943-51