Abstract

The aim of this research program is to understand in molecular detail the mechanism by which the replication of Herpes Simplex-1 (HSV-1) virus DNA is initiated and sustained. We propose to reconstitute in vitro with purified enzymes whose structure and mechanism we will examine in detail, the orderly, semiconservative replication of the HSV-1 genome. We expect that these studies should provide us with an insight into the mechanism by which replication of a eukaryotic genome is initiated. They should also yield information about the replication of a class of animal viruses that are of great public health importance. The investigation will be organized along the following lines: A. Analysis of purified HSV-1 encoded replication enzymes 1. Origin binding protein (UL9 protein) a. Origin specific helicase activity b. Interaction of UL9 protein with other HSV-1 replication enzymes c. Phosphorylation of LTL9 protein 2. DNA polymerase - UL42 protein complex a. Three dimensional structure of DNA polymerase b. Mechanism of DNA polymerase reaction c. 5' -> 3' exonuclease (RNase H) activity of DNA polymerase d. Phosphorylation of UL42 protein 3. DNA helicase - primase 4. Single-strand DNA binding protein - ICP8 B. Reconstitution of Origin (Ori-s) dependent DNA replication 1. Role of origin binding protein in initiation 2. Specific approaches to reconstitution of Ori-s dependent DNA replication a. Transcriptional activation of initiation b. Requirement for host factors c. Mechanistic studies 3. Regulation of HSV-L DNA replication

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI026538-06
Application #
3140288
Study Section
Biochemistry Study Section (BIO)
Project Start
1988-07-01
Project End
1998-06-30
Budget Start
1993-07-01
Budget End
1994-06-30
Support Year
6
Fiscal Year
1993
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

Huang, Ke-Jung; Ku, Chia-Chi; Lehman, I Robert (2006) Endonuclease G: a role for the enzyme in recombination and cellular proliferation. Proc Natl Acad Sci U S A 103:8995-9000
Eom, Chi-Yong; Heo, Won Do; Craske, Madeleine L et al. (2004) The neural F-box protein NFB42 mediates the nuclear export of the herpes simplex virus type 1 replication initiator protein (UL9 protein) after viral infection. Proc Natl Acad Sci U S A 101:4036-40
Eom, Chi-Yong; Lehman, I Robert (2003) Replication-initiator protein (UL9) of the herpes simplex virus 1 binds NFB42 and is degraded via the ubiquitin-proteasome pathway. Proc Natl Acad Sci U S A 100:9803-7
Eom, Chi-Yong; Lehman, I Robert (2002) The human DnaJ protein, hTid-1, enhances binding of a multimer of the herpes simplex virus type 1 UL9 protein to oris, an origin of viral DNA replication. Proc Natl Acad Sci U S A 99:1894-8
He, X; Lehman, I R (2001) An initial ATP-independent step in the unwinding of a herpes simplex virus type I origin of replication by a complex of the viral origin-binding protein and single-strand DNA-binding protein. Proc Natl Acad Sci U S A 98:3024-8
He, X; Lehman, I R (2000) Unwinding of a herpes simplex virus type 1 origin of replication (Ori(S)) by a complex of the viral origin binding protein and the single-stranded DNA binding protein. J Virol 74:5726-8
Lee, S S; Lehman, I R (1999) The interaction of herpes simplex type 1 virus origin-binding protein (UL9 protein) with Box I, the high affinity element of the viral origin of DNA replication. J Biol Chem 274:18613-7
Lehman, I R; Boehmer, P E (1999) Replication of herpes simplex virus DNA. J Biol Chem 274:28059-62
Falkenberg, M; Elias, P; Lehman, I R (1998) The herpes simplex virus type 1 helicase-primase. Analysis of helicase activity. J Biol Chem 273:32154-7
Boehmer, P E; Lehman, I R (1997) Herpes simplex virus DNA replication. Annu Rev Biochem 66:347-84

Showing the most recent 10 out of 39 publications