The adenovirus E1B 55-kDa (55K) protein functions during virus growth and virus-mediated cellular transformation. During virus growth, the 55K protein binds the E4orf6 protein and regulates mRNA biogenesis. During transformation, the 55K protein binds the tumor suppressor p53 and blocks transcription from p53-responsive genes including those that promote apoptosis and induce G1 growth arrest. However, most cells infected with the 55K-mutant virus do not undergo apoptosis, do not arrest in G1, and do not fail to replicate mutant viral DNA. Consequently, the link between 55K function in virus growth and transformation remains unclear. This work will elucidate this link as part of the larger goal of understanding the means by which adenovirus targets mechanisms of cellular growth control for virus growth. Because 55K-mutant viruses grow only in HeLa cells that were infected in early S phase but not in G1, the 55K protein overcomes a restriction imposed on virus growth by the cell cycle. This restriction and the 55K function that overcomes this restriction will be determined by three specific aims. First, the hypothesis that adenovirus must usurp the control of mRNA for cell cycle-independent growth will be evaluated by comparing the growth of related G1-restricted adenovirus mutants and the control of mRNA transport among infected cells that are permissive and restrictive for virus growth. Second, heterokaryons of permissive and restrictive cells will be used to elucidate the cellular basis for this restriction and determine the dominance of the restrictive phenotype. Third, cellular proteins targeted by 55K/E4orf6 complex for cell cycle-independent virus growth will be identified and the cell cycle- regulation of these proteins determined. This study will determine how the 55K protein subverts cell cycle controls for virus growth. This may represent the first link between the roll of the 55K protein in lytic growth and viral transformation. These studies will increase our understanding of the mechanisms of viral pathogenesis and viral oncogenesis as well as fundamental mechanisms of cellular growth control. Furthermore, this study will identify adenovirus mutants that are restricted for growth in S phase cells. Such replication competent viruses can be used as oncolytic agents to treat rapidly growing human tumors.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
3R01CA077342-05S2
Application #
6947659
Study Section
Virology Study Section (VR)
Program Officer
Daschner, Phillip J
Project Start
1999-04-01
Project End
2005-03-31
Budget Start
2003-04-01
Budget End
2005-03-31
Support Year
5
Fiscal Year
2004
Total Cost
$21,525
Indirect Cost
Name
Wake Forest University Health Sciences
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
937727907
City
Winston-Salem
State
NC
Country
United States
Zip Code
27157
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