Abstract

It is proposed to analyze the mechanism of initiation of DNA replication in three closely related small DNA viruses phi X174, G4 and ST-1) that replicate their DNA using different host cell proteins. The structure of the initiation site in both strands of the DNA duplex will be investigated, together with the proteins that each strand requires for initiation of synthesis of its complementary DNA strand. The structure and relationship of the two sites will be correlated with the model of DNA replication that each virus uses and the structure of the initiation sites will be changed by in vivo and in vitro mutagenesis in order to analyze the requirements of structure on their function. These studies will indicate whether the initiation of synthesis of each strand of the DNA duplex is independent. The overall objective is to learn enough about the sites of initiation of DNA synthesis to be able to specifically inactivate different origins of DNA replication in the same cell and to be able to design a rational method of specifically inactivating invading viruses or resident oncogenic viruses.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM038292-12A3
Application #
2179270
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1986-08-01
Project End
1998-03-31
Budget Start
1994-04-01
Budget End
1995-03-31
Support Year
12
Fiscal Year
1994
Total Cost
Indirect Cost

  Institution

Name
New York University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
004514360
City
New York
State
NY
Country
United States
Zip Code
10012

  Publications

Rodina, Anna; Godson, G Nigel (2006) Role of conserved amino acids in the catalytic activity of Escherichia coli primase. J Bacteriol 188:3614-21
Godson, G N; Schoenich, J; Sun, W et al. (2000) Identification of the magnesium ion binding site in the catalytic center of Escherichia coli primase by iron cleavage. Biochemistry 39:332-9
Yajnik, V; Godson, G N (1993) Selective decay of Escherichia coli dnaG messenger RNA is initiated by RNase E. J Biol Chem 268:13253-60
Nesin, M; Svec, P; Lupski, J R et al. (1990) Cloning and nucleotide sequence of a chromosomally encoded tetracycline resistance determinant, tetA(M), from a pathogenic, methicillin-resistant strain of Staphylococcus aureus. Antimicrob Agents Chemother 34:2273-6
Hiasa, H; Sakai, H; Komano, T et al. (1990) Structural features of the priming signal recognized by primase: mutational analysis of the phage G4 origin of complementary DNA strand synthesis. Nucleic Acids Res 18:4825-31
Hiasa, H; Sakai, H; Tanaka, K et al. (1989) Mutational analysis of the primer RNA template region in the replication origin (oric) of bacteriophage G4: priming signal recognition by Escherichia coli primase. Gene 84:9-16
Almond, N; Yajnik, V; Svec, P et al. (1989) An Escherichia coli cis-acting antiterminator sequence: the dnaG nut site. Mol Gen Genet 216:195-203
Hiasa, H; Tanaka, K; Sakai, H et al. (1989) Distinct functional contributions of three potential secondary structures in the phage G4 origin of complementary DNA strand synthesis. Gene 84:17-22
Sakai, H; Hiasa, H; Iwamoto, K et al. (1988) Role of the potential secondary structures in phage G4 origin of complementary DNA strand synthesis. Gene 71:323-30
Nesin, M; Lupski, J R; Godson, G N (1988) Role of the 5' upstream sequence and tandem promoters in regulation of the rpsU-dnaG-rpoD macromolecular synthesis operon. J Bacteriol 170:5759-64