Abstract | BACKGROUND: METHODS:
c-Src kinase inhibitors ( SU6656, PP1, and PP2), adenovectors [wild-type and dominant-negative (DN) c-src] or siRNA (targeting c-src or pyk2) were used to inhibit, compete with or knockdown c-Src in Jurkat C, Jurkat E6-1, Hut 78 or Kit225 T-cell lines. Cells were then infected with HIV-1 luciferase reporter virus expressing VSV-G or HXB2(X4) envelope, and luciferase activity was measured after 2 days. Reverse transcriptase activity and viral cDNA were measured 1 hour after infection, whereas integrated virus was measured 12 hours after infection. RESULTS: Pretreating Jurkat T-cells with SU6656 led to increased VSV-G luciferase activity. In the adenovector experiments, T-cells overexpressing dominant-negative c-Src, but not wild-type c-Src, showed increased luciferase activity after VSV-G infection. siRNA knockdown of c-Src or Pyk2, followed by HXB2 infection in Jurkat T-cells, lead to increased reverse transcriptase activity, viral cDNA, integrated virus, and increased luciferase activity. CONCLUSIONS: Pyk2 is known to interact with c-Src. Thus, Pyk2 activation that coincides with increased c-Src activity during HIV-1 infection could be responsible for c-Src activation. Reduced c-Src activation increases HIV-1 reverse transcription, integration, and/or transcription, suggesting the high c-Src activity seen early in HIV-1 infection may be a cellular response to slow or prevent early infection in CD4 T-cells.
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Authors | Stephen D S McCarthy, Daniel Jung, Darinka Sakac, Donald R Branch |
Journal | Journal of acquired immune deficiency syndromes (1999)
(J Acquir Immune Defic Syndr)
Vol. 66
Issue 2
Pg. 118-26
(Jun 01 2014)
ISSN: 1944-7884 [Electronic] United States |
PMID | 24413044
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- 4-amino-5-(4-methylphenyl)-7-(tert-butyl)pyrazolo(3,4-d)pyrimidine
- AG 1879
- DNA, Viral
- G protein, vesicular stomatitis virus
- Indoles
- Membrane Glycoproteins
- Pyrazoles
- Pyrimidines
- SU 6656
- Sulfonamides
- Viral Envelope Proteins
- CSK Tyrosine-Protein Kinase
- Focal Adhesion Kinase 2
- PTK2B protein, human
- src-Family Kinases
- CSK protein, human
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Topics |
- CD4-Positive T-Lymphocytes
(virology)
- CSK Tyrosine-Protein Kinase
- Cell Line, Tumor
- DNA, Viral
(isolation & purification)
- Focal Adhesion Kinase 2
(antagonists & inhibitors, genetics, metabolism)
- HIV Infections
(metabolism)
- HIV-1
(physiology)
- Humans
- Indoles
(pharmacology)
- Membrane Glycoproteins
(metabolism)
- Phosphorylation
- Pyrazoles
(pharmacology)
- Pyrimidines
(pharmacology)
- Sulfonamides
(pharmacology)
- Transduction, Genetic
- Viral Envelope Proteins
(metabolism)
- src-Family Kinases
(antagonists & inhibitors, genetics, metabolism)
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