A molecular study of chromatin containing integrated murine leukemia virus DNA will be performed. These experiments will provide model systems to investigate 1) chromatin configuration around gene regulatory regions and 2) chromosomal position effect. Moloney murine leukemia viruses (M-MuLV's) containing altered LTR's will be generated by recombinant DNA manipulations. DNA sequences conferring hormone or metal responsiveness will be inserted, as will heterologous transcriptional enhancersers. M-MuLV's carrying the modified LTR's will be recovered by transfection, and tested for hormone or metal responsiveness or change in host range. The chromatin structure of the altered proviruses will be investigated, and the appearance of new DNAse I hypersensitive sites or bound proteins will be correlated with the biological activity. Promoter activity of the altered LTR's will also be assessed by transient expression assays. Chromatin structure of transcribed and non-transcribed M-MuLV proviruses from a cell line chronically infected with M-MuLV will be investigated. Recombinant clones of individual M-MuLV proviruses will be used to identify these proviruses. Nuclear run-off transcription will be employed. The chromatin state of the unoccupied host site as well as the proviral integration will be investigated by DNase I digestion. Cellular proteins which associate with the M-MuLV LTR will be identified by procedures which reveal specific DNA-binding proteins. The location of protein binding in the LTR will be mapped, and LTR's which contain deletions and insertions will be studied. Extracts from different cell types will be tested. Correlation with the in vivo chromatin studies may reveal specific proteins responsible for particular features of chromatin structure.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA032454-05
Application #
3170364
Study Section
Virology Study Section (VR)
Project Start
1981-09-01
Project End
1990-04-30
Budget Start
1985-09-01
Budget End
1986-04-30
Support Year
5
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of California Irvine
Department
Type
Schools of Arts and Sciences
DUNS #
161202122
City
Irvine
State
CA
Country
United States
Zip Code
92697
Feuer, G; Fan, H (1990) Substitution of murine transthyretin (prealbumin) regulatory sequences into the Moloney murine leukemia virus long terminal repeat yields infectious virus with altered biological properties. J Virol 64:6130-40
Feuer, G; Taketo, M; Hanecak, R C et al. (1989) Two blocks in Moloney murine leukemia virus expression in undifferentiated F9 embryonal carcinoma cells as determined by transient expression assays. J Virol 63:2317-24
Kitado, H; Fan, H (1989) Chromatin structure of recombinant Moloney murine leukemia virus proviral DNAs that contain tax-responsive sequences from human T-cell lymphotropic virus type II in the presence and absence of tax. J Virol 63:3072-9
Thompson, T; Fan, H (1988) Chromatin structure of hormone-responsive Moloney murine leukemia virus proviruses that contain sequences from mouse mammary tumor virus. Virus Genes 2:83-98
Hanecak, R; Pattengale, P K; Fan, H (1988) Addition of substitution of simian virus 40 enhancer sequences into the Moloney murine leukemia virus (M-MuLV) long terminal repeat yields infectious M-MuLV with altered biological properties. J Virol 62:2427-36
Kitado, H; Chen, I S; Shah, N P et al. (1987) U3 sequences from HTLV-I and -II LTRs confer pX protein response to a murine leukemia virus LTR. Science 235:901-4
Fan, H; Mittal, S; Chute, H et al. (1986) Rearrangements and insertions in the Moloney murine leukemia virus long terminal repeat alter biological properties in vivo and in vitro. J Virol 60:204-14
Thompson, T; Fan, H (1985) Mapping of DNase I-hypersensitive sites in the 5' and 3' long terminal repeats of integrated moloney murine leukemia virus proviral DNA. Mol Cell Biol 5:601-9
Overhauser, J; Fan, H (1985) Generation of glucocorticoid-responsive Moloney murine leukemia virus by insertion of regulatory sequences from murine mammary tumor virus into the long terminal repeat. J Virol 54:133-44
Davis, B; Linney, E; Fan, H (1985) Suppression of leukaemia virus pathogenicity by polyoma virus enhancers. Nature 314:550-3