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Originally published In Press as doi:10.1074/jbc.M405761200 on June 20, 2004

J. Biol. Chem., Vol. 279, Issue 34, 35822-35828, August 20, 2004
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Human Apolipoprotein B mRNA-editing Enzyme-catalytic Polypeptide-like 3G (APOBEC3G) Is Incorporated into HIV-1 Virions through Interactions with Viral and Nonviral RNAs*

Evguenia S. Svarovskaia{ddagger}, Hongzhan Xu{ddagger}, Jean L. Mbisa{ddagger}, Rebekah Barr{ddagger}, Robert J. Gorelick§, Akira Ono{ddagger}, Eric O. Freed{ddagger}, Wei-Shau Hu{ddagger}, and Vinay K. Pathak{ddagger}

From the {ddagger}HIV Drug Resistance Program and §AIDS Vaccine Program, Science Applications International Corporation-Frederick, Inc., NCI-Frederick, National Institutes of Health, Frederick, Maryland 21702

Apolipoprotein B mRNA-editing enzyme-catalytic polypeptide-like 3G (APOBEC3G) is a host cytidine deaminase that is packaged into virions and confers resistance to retroviral infection. APOBEC3G deaminates deoxycytidines in minus strand DNA to deoxyuridines, resulting in G to A hypermutation and viral inactivation. Human immunodeficiency virus type 1 (HIV-1) virion infectivity factor counteracts the antiviral activity of APOBEC3G by inducing its proteosomal degradation and preventing virion incorporation. To elucidate the mechanism of viral suppression by APOBEC3G, we developed a sensitive cytidine deamination assay and analyzed APOBEC3G virion incorporation in a series of HIV-1 deletion mutants. Virus-like particles derived from constructs in which pol, env, and most of gag were deleted still contained high levels of cytidine deaminase activity; in addition, coimmunoprecipitation of APOBEC3G and HIV-1 Gag in the presence and absence of RNase A indicated that the two proteins do not interact directly but form an RNase-sensitive complex. Viral particles lacking HIV-1 genomic RNA which were generated from the gag-pol expression constructs pC-Help and pSYNGP packaged APOBEC3G at 30–40% of the wild-type level, indicating that interactions with viral RNA are not necessary for incorporation. In addition, viral particles produced from an nucleocapsid zinc finger mutant contained ~1% of the viral genomic RNA but ~30% of the cytidine deaminase activity. The reduction in APOBEC3G incorporation was equivalent to the reduction in the total RNA present in the nucleocapsid mutant virions. These results indicate that interactions with viral proteins or viral genomic RNA are not essential for APOBEC3G incorporation and suggest that APOBEC3G interactions with viral and nonviral RNAs that are packaged into viral particles are sufficient for APOBEC3G virion incorporation.

Received for publication, May 24, 2004

* This work was supported in part with Federal funds from the NCI, National Institutes of Health Contract NO1-CO-12400 with Science Applications International Corporation-Frederick, Inc. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondance should be addressed: HIV Drug Resistance Program, Center for Cancer Research, NCI-Frederick, P. O. Box B, Bldg. 535, Rm. 334, Frederick, MD 21702. Fax: 301-846-6013; E-mail: vpathak{at}ncifcrf.gov.


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