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Journal of Virology, January 2005, p. 910-917, Vol. 79, No. 2
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.2.910-917.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Covalent Modification of Human Immunodeficiency Virus Type 1 p6 by SUMO-1

Cagan Gurer,1 Lionel Berthoux,1 and Jeremy Luban1,2*

Departments of Microbiology,1 Medicine, Columbia University, New York, New York2

Received 22 June 2004/ Accepted 3 August 2004

The p6 domain of the human immunodeficiency virus type 1 (HIV-1) Gag polyprotein mediates virion budding from infected cells via protein-protein contacts with the class E vacuolar protein sorting factors, Tsg101 and AIP1/ALIX. Interaction with Tsg101 is strengthened by covalent attachment of monovalent ubiquitin to HIV-1 p6. To identify additional host factors that bind to HIV-1 p6, a human cDNA library was screened in the yeast two-hybrid system. HIV-1 p6 was found to interact with small ubiquitin-like modifier 1 (SUMO-1) as well as the E2 SUMO-1 transfer enzyme, Ubc9. Interaction with p6 was also detected with Daxx, a cellular protein to which SUMO-1 is sometimes covalently attached. SUMO-1 was incorporated into HIV-1 virions where it was protected within the virion membrane from digestion by exogenous protease. Of the two lysine residues in p6, lysine 27 uniquely served as a site of covalent SUMO-1 attachment. As previously reported, though, HIV-1 bearing the p6-K27R mutation replicated just like the wild type. Overproduction of SUMO-1 in HIV-1 producer cells had no apparent effect on virion release or on virion protein or RNA content. Infectivity of the resulting virions, though, was decreased, with the defect occurring after membrane fusion, at the time of viral cDNA synthesis. HIV-1 bearing the p6-K27R mutation was insensitive to SUMO-1 overexpression, suggesting that covalent attachment of SUMO-1 to p6 is detrimental to HIV-1 replication.

* Corresponding author. Mailing address: Microbiology Department, Columbia University, 701 W. 168th St., New York, NY 10032. Phone: (212) 305-8710. Fax: (212) 305-0333. E-mail: jl45{at}columbia.edu.

Journal of Virology, January 2005, p. 910-917, Vol. 79, No. 2
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.2.910-917.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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