Abstract

The aim of this research program is to understand in molecular detail the mechanism of DNA replication in a model eukaryote, Drosophila melanogaster. The investigation will be organized along the following lines. 1. DNA chain elongation: a. Dissociation and reconstitution of Drosophila polymerase-primase. (1) The 182 and 73 kDa subunits (2) The 60 and 50 kDa primase subunits (3) A cryptic 31-51 exonuclease in mammalian and yeast polymerase-primase (4) Use of Drosophila 182 kDa subunit to identify Drosophila single-stranded DNA binding protein (5) Replication of chromatin by polymerase-primase b.DNA polymerase 6 in Drosophila embryos 2. Initiation of DNA replication at a Drosophila origin in vitro a. Identification of a chorion origin binding protein(s) b. Initiation in vitro of chorion DNA replication 3. Regulation of DNA replication a. Changes in polymerase-primase during embryogenesis

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37GM006196-33
Application #
3484057
Study Section
Physiological Chemistry Study Section (PC)
Project Start
1974-09-01
Project End
1994-08-31
Budget Start
1991-09-01
Budget End
1992-08-31
Support Year
33
Fiscal Year
1991
Total Cost
Indirect Cost

  Institution

Name
Stanford University
Department
Type
Schools of Medicine
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305

  Publications

Mitsis, P G; Kowalczykowski, S C; Lehman, I R (1993) A single-stranded DNA binding protein from Drosophila melanogaster: characterization of the heterotrimeric protein and its interaction with single-stranded DNA. Biochemistry 32:5257-66
Chiang, C S; Mitsis, P G; Lehman, I R (1993) DNA polymerase delta from embryos of Drosophila melanogaster. Proc Natl Acad Sci U S A 90:9105-9
Crute, J J; Tsurumi, T; Zhu, L A et al. (1989) Herpes simplex virus 1 helicase-primase: a complex of three herpes-encoded gene products. Proc Natl Acad Sci U S A 86:2186-9
Lehman, I R; Kaguni, L S (1989) DNA polymerase alpha. J Biol Chem 264:4265-8
Reyland, M E; Lehman, I R; Loeb, L A (1988) Specificity of proofreading by the 3'----5' exonuclease of the DNA polymerase-primase of Drosophila melanogaster. J Biol Chem 263:6518-24
Kaguni, L S; Lehman, I R (1988) Eukaryotic DNA polymerase-primase: structure, mechanism and function. Biochim Biophys Acta 950:87-101
Crute, J J; Mocarski, E S; Lehman, I R (1988) A DNA helicase induced by herpes simplex virus type 1. Nucleic Acids Res 16:6585-96
Elias, P; Lehman, I R (1988) Interaction of origin binding protein with an origin of replication of herpes simplex virus 1. Proc Natl Acad Sci U S A 85:2959-63
Cotterill, S M; Reyland, M E; Loeb, L A et al. (1987) A cryptic proofreading 3'----5' exonuclease associated with the polymerase subunit of the DNA polymerase-primase from Drosophila melanogaster. Proc Natl Acad Sci U S A 84:5635-9
Cox, M M; Lehman, I R (1987) Enzymes of general recombination. Annu Rev Biochem 56:229-62

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