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Purification of Viral DNA for the Identification of Associated Viral and Cellular Proteins
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Role of cellular phosphatase cdc25C in herpes simplex virus 1 replication
Benjamin A Smith-Donald1, Lizette O Durand, Bernard Roizman
1The University of Chicago, Marjorie B. Kovler Viral Oncology Laboratories, 910 East 58th St., Chicago, IL 60637. bernard.roizman@bsd.uchicago.edu.
Journal of Virology
|February 15, 2008
Summary
Herpes simplex virus 1 infection involves ICP22, which upregulates gamma(2) proteins. This study shows the mitotic phosphatase cdc25C plays a role in viral replication beyond activating cdc2.
Area of Science:
- Virology
- Molecular Biology
- Cell Cycle Regulation
Background:
- Herpes simplex virus 1 (HSV-1) infection involves the viral protein ICP22.
- ICP22 upregulates specific gamma(2) viral proteins through a complex pathway.
- This pathway involves cell cycle regulators like cyclins, cdc2, and the phosphatase cdc25C.
Purpose of the Study:
- To investigate the role of the mitotic phosphatase cdc25C in the ICP22-dependent pathway of gamma(2) gene upregulation during HSV-1 infection.
- To determine if cdc25C's function extends beyond cdc2 activation in this viral process.
Main Methods:
- Utilized cdc25C knockout (cdc25C(-/-)) and wild-type (cdc25C(+/+)) mouse embryonic fibroblast cells.
- Infected these cells with HSV-1 (wild-type and ICP22-deficient strains).
- Analyzed protein levels (cyclin B1, cdc2, U(S)11, U(L)38, topoisomerase IIalpha) and infectious virus production.
Main Results:
- Cyclin B1 degradation rates were similar in cdc25C(+/+) and cdc25C(-/-) cells.
- cdc2 levels increased in both cell types.
- Accumulation of viral proteins U(S)11 and U(L)38, and infectious virus yield were reduced in cdc25C(-/-) cells compared to wild-type.
- This reduction was more pronounced in cdc25C(-/-) cells infected with ICP22-deficient HSV-1.
Conclusions:
- The phosphatase cdc25C plays a significant role in HSV-1 replication.
- The function of cdc25C in viral replication is not limited to its role in activating cdc2 for the ICP22-dependent cascade.
- cdc25C influences the upregulation of gamma(2) gene expression during HSV-1 infection through mechanisms beyond cdc2 activation.
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