Abstract

The goal of the proposed research is to identify and characterize the proteins involved in the replication of the genome of the DNA tumor virus SV40. We expect that much of what is learned about the replication of this simple viral chromosome will be relevant to understanding cellular DNA replication and its control. During the past grant period we achieved the reconstitution of origin-dependent DNA replication in vitro with purified proteins and characterized the biochemical mechanisms of initiation and elongation of nascent DNA chains. We also obtained cDNA clones corresponding to the cellular proteins involved in the initiation of SV40 DNA synthesis. During the next grant period we intend to make use of these reagents to analyze the functional roles of the individual cellular proteins in the initiation pathway and to characterize the critical interactions that determine the specificity and host range of viral DNA replication. We also plan to make use of the information obtained in these (and previous) studies to seek cellular counterparts of SV40 T antigen.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37CA040414-14
Application #
2700375
Study Section
Experimental Virology Study Section (EVR)
Program Officer
Wong, May
Project Start
1985-07-01
Project End
2000-04-30
Budget Start
1998-05-01
Budget End
1999-04-30
Support Year
14
Fiscal Year
1998
Total Cost
Indirect Cost

  Institution

Name
Johns Hopkins University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218

  Publications

Dai, Jianli; Chuang, Ray-Yuan; Kelly, Thomas J (2005) DNA replication origins in the Schizosaccharomyces pombe genome. Proc Natl Acad Sci U S A 102:337-42
Vashee, S; Simancek, P; Challberg, M D et al. (2001) Assembly of the human origin recognition complex. J Biol Chem 276:26666-73
Moarefi, I F; Small, D; Gilbert, I et al. (1993) Mutation of the cyclin-dependent kinase phosphorylation site in simian virus 40 (SV40) large T antigen specifically blocks SV40 origin DNA unwinding. J Virol 67:4992-5002
Virshup, D M; Russo, A A; Kelly, T J (1992) Mechanism of activation of simian virus 40 DNA replication by protein phosphatase 2A. Mol Cell Biol 12:4883-95
Randall, S K; Kelly, T J (1992) The fate of parental nucleosomes during SV40 DNA replication. J Biol Chem 267:14259-65
Erdile, L F; Collins, K L; Russo, A et al. (1991) Initiation of SV40 DNA replication: mechanism and control. Cold Spring Harb Symp Quant Biol 56:303-13
Scheidtmann, K H; Virshup, D M; Kelly, T J (1991) Protein phosphatase 2A dephosphorylates simian virus 40 large T antigen specifically at residues involved in regulation of DNA-binding activity. J Virol 65:2098-101
Weinberg, D H; Collins, K L; Simancek, P et al. (1990) Reconstitution of simian virus 40 DNA replication with purified proteins. Proc Natl Acad Sci U S A 87:8692-6
Virshup, D M; Kauffman, M G; Kelly, T J (1989) Activation of SV40 DNA replication in vitro by cellular protein phosphatase 2A. EMBO J 8:3891-8
Cheng, L; Kelly, T J (1989) Transcriptional activator nuclear factor I stimulates the replication of SV40 minichromosomes in vivo and in vitro. Cell 59:541-51

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