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c-Myb influences HIV type 1 gene expression and virus production
M J Churchill1, R G Ramsay, D I Rhodes
1AIDS Molecular Biology Unit, National Centre in HIV Virology Research, Macfarlane Burnet Centre for Medical Research, Fairfield, Victoria 3078, Australia. churchil@burnet.edu.au
AIDS Research and Human Retroviruses
|November 16, 2001
Summary
The transcription factor c-Myb regulates human immunodeficiency virus type 1 (HIV-1) replication in T cells. Inhibiting c-Myb significantly reduces HIV-1 gene expression and virus production.
Area of Science:
- Molecular Biology
- Virology
- Immunology
Background:
- The c-Myb proto-oncogene is expressed in proliferating T lymphocytes.
- The human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR) contains multiple c-Myb binding sites, suggesting a regulatory role for c-Myb in HIV-1 gene expression and replication.
Purpose of the Study:
- To investigate the role of c-Myb in regulating HIV-1 gene expression and replication in CD4(+) T cells.
Main Methods:
- Transient transfection of CEM cells to increase c-Myb levels and assess HIV-1 LTR-driven gene expression.
- Mutation of a high-affinity Myb-binding site in the HIV-1 LTR.
- Treatment of MT-2 cells with c-myb antisense oligonucleotides to inhibit c-Myb expression.
- Measurement of HIV-1 replication using reverse transcriptase activity and cytopathic effects.
Main Results:
- Increasing cellular c-Myb levels led to a 10- to 20-fold activation of HIV-1 LTR-driven gene expression.
- Mutation of a key c-Myb binding site reduced this activation by 60-70%.
- Inhibition of c-Myb expression via antisense oligonucleotides decreased HIV-1 replication by 85%.
Conclusions:
- c-Myb expression significantly affects HIV-1 replication in CD4(+) T cells.
- The regulatory effect of c-Myb on HIV-1 is mediated through the HIV-1 LTR and is independent of cell proliferation.