Sequence variation plays a significant role in the pathogenesis and persistence of infections and is a major obstacle in the development of vaccines as well as therapies against lethal diseases caused by retroviruses. It is the goal of the investigator to further elucidate the basic molecular mechanisms of retroviral variation. More specifically, she plans to focus on the mechanisms of retroviral recombination. It is the investigator's hypothesis that certain nucleic acid sequences are more prone to recombination than others and that at least some of these potential recombination hotspots correlate with pause and termination sites for reverse transcriptase. In order to test this hypothesis, she will: 1) determine whether hotspots for mutation correlate with hotspots for retroviral recombination; 2) determine the pause and termination sites for reverse transcriptase in vivo; and 3) identify naturally occurring retroviral sequences which serve as hotspots for recombination. The basic in vivo system will utilize retroviral vectors and helper cells that are designed to isolate one cycle of viral replication. Recombinants will be analyzed by a variety of methods including restriction enzyme mapping, base mutation screening, and DNA sequencing. For the pause and termination assay, unintegrated viral DNA will be analyzed by polymerase chain reaction (PCR). Study of these basic mechanisms will contribute to understanding the role of retroviral strand transfer.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29CA072239-04
Application #
2895724
Study Section
AIDS and Related Research Study Section 3 (ARRC)
Program Officer
Cole, John S
Project Start
1996-07-01
Project End
2001-04-30
Budget Start
1999-05-01
Budget End
2000-04-30
Support Year
4
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Wright State University
Department
Other Basic Sciences
Type
Schools of Medicine
DUNS #
City
Dayton
State
OH
Country
United States
Zip Code
45435
Huang, K J; Wooley, D P (2005) A new cell-based assay for measuring the forward mutation rate of HIV-1. J Virol Methods 124:95-104
Taylor, R J; Schols, D; Wooley, D P (2004) Restricted entry of R5 HIV Type 1 strains into eosinophilic cells. AIDS Res Hum Retroviruses 20:1244-53
Ponferrada, V G; Mauck, B S; Wooley, D P (2003) The envelope glycoprotein of human endogenous retrovirus HERV-W induces cellular resistance to spleen necrosis virus. Arch Virol 148:659-75
Bircher, L A; Rigano, J C; Ponferrada, V G et al. (2002) High fidelity of homologous retroviral recombination in cell culture. Arch Virol 147:1665-83
Huang, Kelly J; Alter, Gerald M; Wooley, Dawn P (2002) The reverse transcriptase sequence of human immunodeficiency virus type 1 is under positive evolutionary selection within the central nervous system. J Neurovirol 8:281-94
Wooley, D P; Peterson, K T; Taylor, R J et al. (2000) Strain-dependent productive infection of a unique eosinophilic cell line by human immunodeficiency virus type 1. AIDS Res Hum Retroviruses 16:1405-15
Wooley, D P; Bircher, L A; Smith, R A (1998) Retroviral recombination is nonrandom and sequence dependent. Virology 243:229-34
Wooley, D P; Smith, R A; Czajak, S et al. (1997) Direct demonstration of retroviral recombination in a rhesus monkey. J Virol 71:9650-3