Abstract

We are studying replication of parvovirus H-1 DNA in vitro as a model for mammalian DNA replication. An activity from uninfected cells capable of converting viral single stranded DNA to double stranded replicative form has been purified and identified as DNA polymerase gamma. Additional stimulatory factors for this reaction will be purified and characterized, and the studies will be extended to the replication of double stranded replicative form. We have shown that methotrexate and FdUrd cause expansion of the pools of dUTP (and FdUTP) and lead to incorporation of dUMP (and FdUMP) into DNA. Further studies are examining the relationship of these effects to DNA fragmentation and cell death.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA011705-20
Application #
3163543
Study Section
Physiological Chemistry Study Section (PC)
Project Start
1975-01-01
Project End
1991-03-31
Budget Start
1989-07-01
Budget End
1991-03-31
Support Year
20
Fiscal Year
1989
Total Cost
Indirect Cost

  Institution

Name
University of California San Diego
Department
Type
Schools of Medicine
DUNS #
077758407
City
La Jolla
State
CA
Country
United States
Zip Code
92093

  Publications

Goulian, M; Heard, C J (1991) Discrimination between mammalian RNases H-1 and H-2. Anal Biochem 192:398-402
Goulian, M; Heard, C J; Grimm, S L (1990) Purification and properties of an accessory protein for DNA polymerase alpha/primase. J Biol Chem 265:13221-30
Goulian, M; Richards, S H; Heard, C J et al. (1990) Discontinuous DNA synthesis by purified mammalian proteins. J Biol Chem 265:18461-71
Goulian, M; Herrmann, S M; Sackett, J W et al. (1990) Two forms of DNA polymerase delta from mouse cells. Purification and properties. J Biol Chem 265:16402-11
Goulian, M; Grimm, S L (1990) Three cytoplasmic DNA polymerases that utilize poly(rA).oligo(dT). Biochem Biophys Res Commun 170:627-34
Goulian, M; Heard, C J (1990) The mechanism of action of an accessory protein for DNA polymerase alpha/primase. J Biol Chem 265:13231-9
Goulian, M; Heard, C J (1990) An inhibitor of DNA polymerases alpha and delta in calf thymus DNA. Nucleic Acids Res 18:4791-6
Goulian, M; Heard, C J (1989) Intact DNA polymerase alpha/primase from mouse cells. Purification and structure. J Biol Chem 264:19407-15
Goulian, M; Bleile, B M; Dickey, L M et al. (1986) Mechanism of thymineless death. Adv Exp Med Biol 195 Pt B:89-95
Ingraham, H A; Dickey, L; Goulian, M (1986) DNA fragmentation and cytotoxicity from increased cellular deoxyuridylate. Biochemistry 25:3225-30