We have isolated a large collection of temperature-sensitive (ts) mutants in established Chinese Hamster cell lines (CHO, V79) and initiated their characterization. Approximately 5% are phenotypically ts DNA-. These will be further tested for the effect of the ts cell function for adenovirus DNA synthesis in vivo and in vitro so as to define biochemically the defect. A previously isolated class of ts DNA- mutants in mouse 3T3 has been shown to be restrictive for polyomaviral DS in vivo and in vitro. A biochemical defect has been identified in DNA polymerase (polAlpha2). These mutants and selected CHO mutants will be further utilized for isolation of the wild-type homolog of the mutant gene (by gene transfer and recombinant DNA methodologies) and subsequent definition of the mutant polypeptide. Finally, we will determine whether SV40 DNA synthesis can be positively effected in CHO by increasing levels of viral T antigen, viral origins of DNA replication, or both. If successful, such manipulations will be used to extend the utility of the CHO mutants to virus-cell interactions with SV40 as well, permitting the definition of additional cellular functions. Together, these studies should significantly increase our information on viral and cell DNA replication, a fundamental aspect of carcinogenesis.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
3R01CA023002-13S1
Application #
3165986
Study Section
Experimental Virology Study Section (EVR)
Project Start
1977-06-01
Project End
1990-11-30
Budget Start
1988-12-01
Budget End
1990-11-30
Support Year
13
Fiscal Year
1990
Total Cost
Indirect Cost
Name
University of Medicine & Dentistry of NJ
Department
Type
Schools of Medicine
DUNS #
605799469
City
Newark
State
NJ
Country
United States
Zip Code
07107
Chowdary, D R; Dermody, J J; Jha, K K et al. (1994) Accumulation of p53 in a mutant cell line defective in the ubiquitin pathway. Mol Cell Biol 14:1997-2003
Wu, Y; Hickey, R; Lawlor, K et al. (1994) A 17S multiprotein form of murine cell DNA polymerase mediates polyomavirus DNA replication in vitro. J Cell Biochem 54:32-46
Wojcik, B E; Dermody, J J; Ozer, H L et al. (1990) Temperature-sensitive DNA mutant of Chinese hamster ovary cells with a thermolabile ribonucleotide reductase activity. Mol Cell Biol 10:5688-99
Radna, R L; Foellmer, B; Feldman, L A et al. (1987) Restriction of human adenovirus replication in Chinese hamster cell lines and their hybrids with human cells. Virus Res 8:277-99
Dermody, J J; Wojcik, B E; Du, H et al. (1986) Identification of temperature-sensitive DNA- mutants of Chinese hamster cells affected in cellular and viral DNA synthesis. Mol Cell Biol 6:4594-601
Zeng, G C; Zannis-Hadjopoulos, M; Ozer, H L et al. (1985) Defective DNA topoisomerase I activity in a DNAts mutant of Balb/3T3 cells. Somat Cell Mol Genet 11:557-69
Zeng, G C; Ozer, H L; Hand, R (1985) Further characterization of the phenotype of ts20, a DNAts mutant of BALB/3T3 cells. Exp Cell Res 160:184-96
La Bella, F; Ozer, H L (1985) Differential replication of SV40 and polyoma DNAs in Chinese hamster ovary cells. Virus Res 2:329-44