HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 280, Issue 9, 8387-8396, March 4, 2005

This Article Full Text Full Text (PDF) All Versions of this Article: 280/9/8387    most recent M408048200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Wichroski, M. J. Articles by Rana, T. M. Search for Related Content PubMed PubMed Citation Articles by Wichroski, M. J. Articles by Rana, T. M. Social Bookmarking                      What's this?

Analysis of HIV-1 Viral Infectivity Factor-mediated Proteasome-dependent Depletion of APOBEC3G

CORRELATING FUNCTION AND SUBCELLULAR LOCALIZATION^*

Michael J. Wichroski, Kozi Ichiyama, and Tariq M. Rana

From the Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts 01605

To study how HIV-1 viral infectivity factor (Vif) mediates proteasome-dependent depletion of host factor APOBEC3G, functional and nonfunctional Vif-APOBEC3G interactions were correlated with subcellular localization. APOBEC3G localized throughout the cytoplasm and co-localized with -tubulin, 20 S proteasome subunit, and ubiquitin at punctate cytoplasmic bodies that can be used to monitor the Vif-APOBEC3G interaction in the cell. Through immunostaining and live imaging, we showed that a substantial fraction of Vif localized to the nucleus, and this localization was impaired by deletion of amino acids 12-23. When co-expressed, Vif exhibited more pronounced localization to the cytoplasm and reduced the total cellular levels of APOBEC3G but rarely co-localized with APOBEC3G at cytoplasmic bodies. On the contrary, Vif^C114S, which is inactive but continues to interact with APOBEC3G, stably associated with APOBEC3G in the cytoplasm, resulting in complete co-localization at cytoplasmic bodies and a dose-dependent exclusion of Vif^C114S from the nucleus. Following proteasome inhibition, cytoplasmic APOBEC3G levels increased, and both proteins co-accumulated nonspecifically into a vimentin-encaged aggresome. Furthermore in the presence or absence of APOBEC3G, Vif localization was significantly altered by proteasome inhibition, suggesting that aberrant localization may also contribute to the loss of Vif function. Finally mutations at Vif Ile⁹ disrupted the ability of Vif or Vif^C114S to coimmunoprecipitate and to co-localize with APOBEC3G, suggesting that the N terminus of Vif mediates interactions with APOBEC3G. Taken together, these results demonstrate that cytoplasmic Vif-APOBEC3G interactions are required but are not sufficient for Vif to modulate APOBEC3G and can be monitored by co-localization in vivo.

Received for publication, July 16, 2004 , and in revised form, October 14, 2004.

^* This work was supported in part by National Institutes of Health Grants AI 43198 and AI 41404 (to T.M.R.). 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 correspondence should be addressed: Chemical Biology Program, Dept. of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 364 Plantation St., Worcester, MA 01605. Tel.: 508-856-6216; Fax: 508-856-6696; E-mail: tariq.rana{at}umassmed.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				J. L. DeHart, A. Bosque, R. S. Harris, and V. Planelles Human Immunodeficiency Virus Type 1 Vif Induces Cell Cycle Delay via Recruitment of the Same E3 Ubiquitin Ligase Complex That Targets APOBEC3 Proteins for Degradation J. Virol., September 15, 2008; 82(18): 9265 - 9272. [Abstract] [Full Text] [PDF]

				R. P. Bennett, V. Presnyak, J. E. Wedekind, and H. C. Smith Nuclear Exclusion of the HIV-1 Host Defense Factor APOBEC3G Requires a Novel Cytoplasmic Retention Signal and Is Not Dependent on RNA Binding J. Biol. Chem., March 21, 2008; 283(12): 7320 - 7327. [Abstract] [Full Text] [PDF]

				A. Mehle, H. Wilson, C. Zhang, A. J. Brazier, M. McPike, E. Pery, and D. Gabuzda Identification of an APOBEC3G Binding Site in Human Immunodeficiency Virus Type 1 Vif and Inhibitors of Vif-APOBEC3G Binding J. Virol., December 1, 2007; 81(23): 13235 - 13241. [Abstract] [Full Text] [PDF]

				S. Opi, S. Kao, R. Goila-Gaur, M. A. Khan, E. Miyagi, H. Takeuchi, and K. Strebel Human Immunodeficiency Virus Type 1 Vif Inhibits Packaging and Antiviral Activity of a Degradation-Resistant APOBEC3G Variant J. Virol., August 1, 2007; 81(15): 8236 - 8246. [Abstract] [Full Text] [PDF]

				A. Burnett and P. Spearman APOBEC3G Multimers Are Recruited to the Plasma Membrane for Packaging into Human Immunodeficiency Virus Type 1 Virus-Like Particles in an RNA-Dependent Process Requiring the NC Basic Linker J. Virol., May 15, 2007; 81(10): 5000 - 5013. [Abstract] [Full Text] [PDF]

				M. Topper, Y. Luo, M. Zhadina, K. Mohammed, L. Smith, and M. A. Muesing Posttranslational Acetylation of the Human Immunodeficiency Virus Type 1 Integrase Carboxyl-Terminal Domain Is Dispensable for Viral Replication J. Virol., March 15, 2007; 81(6): 3012 - 3017. [Abstract] [Full Text] [PDF]

				S. Gallois-Montbrun, B. Kramer, C. M. Swanson, H. Byers, S. Lynham, M. Ward, and M. H. Malim Antiviral Protein APOBEC3G Localizes to Ribonucleoprotein Complexes Found in P Bodies and Stress Granules J. Virol., March 1, 2007; 81(5): 2165 - 2178. [Abstract] [Full Text] [PDF]

				C. Pace, J. Keller, D. Nolan, I. James, S. Gaudieri, C. Moore, and S. Mallal Population Level Analysis of Human Immunodeficiency Virus Type 1 Hypermutation and Its Relationship with APOBEC3G and vif Genetic Variation. J. Virol., September 1, 2006; 80(18): 9259 - 9269. [Abstract] [Full Text] [PDF]

				Y.-L. Chiu and W. C. Greene APOBEC3 Cytidine Deaminases: Distinct Antiviral Actions along the Retroviral Life Cycle J. Biol. Chem., March 31, 2006; 281(13): 8309 - 8312. [Abstract] [Full Text] [PDF]

				M. Santa-Marta, F. A. da Silva, A. M. Fonseca, and J. Goncalves HIV-1 Vif Can Directly Inhibit Apolipoprotein B mRNA-editing Enzyme Catalytic Polypeptide-like 3G-mediated Cytidine Deamination by Using a Single Amino Acid Interaction and Without Protein Degradation J. Biol. Chem., March 11, 2005; 280(10): 8765 - 8775. [Abstract] [Full Text] [PDF]