Generation and characterization of APOBEC3G-positive 293T cells for HIV-1 Vif study.
We have established a number of 293T cell lines that express a human anti HIV-1 factor APOBEC3G. Out of seven cell clones examined, four were readily demonstrated to express APOBEC3G by immunoblotting analysis. In particular, two clones (A3G-C1 and -C4) were found to produce a much higher level of functional APOBEC3G relative to that by pooled cell clones. The transfection efficiency of all these cell clones were similar to that of the parental cells, producing a comparable level of virions upon transfection of wild type and vif-minus proviral DNA clones. Furthermore, the expression level of APOBEC3G in the best cell line (A3G-C1) was far much higher than those of an APOBEC3G-positive lymphocyte cell line and peripheral blood mononuclear cells. We finally monitored the incorporation of APOBEC3G into virions produced in A3G-C1. APOBEC3G was easily detected in progeny viral particles upon transfection of vif-minus proviral clone but not of wild type. These results indicated that our new A3G-C1 cell line is eminently useful for various studies on the interaction of human APOBEC3G and HIV-1 Vif.
Piroozmand
Ahmad
A
Department of Virology, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima, Japan.
Yamamoto
Yoshihiko
Y
Khamsri
Boonruang
B
Fujita
Mikako
M
Uchiyama
Tsuneo
T
Adachi
Akio
A
D000071480:APOBEC-3G Deaminase; D002460:Cell Line; D003564:Cytidine Deaminase; D016341:Genes, vif / Q000502:physiology*; D015497:HIV-1 / Q000235:genetics / Q000254:growth & development*; D006801:Humans; D009700:Nucleoside Deaminases / Q000032:analysis* / Q000502:physiology; D012097:Repressor Proteins / Q000032:analysis* / Q000502:physiology; D014162:Transfection; D014771:Virion / Q000254:growth & development
D012097:Repressor Proteins; D009700:Nucleoside Deaminases; D000071480:APOBEC-3G Deaminase; C464924:APOBEC3G protein, human; D003564:Cytidine Deaminase