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Journal of Virology, December 2007, p. 12899-12910, Vol. 81, No. 23
0022-538X/07/$08.00+0     doi:10.1128/JVI.01280-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Myristoylation Is Required for Human Immunodeficiency Virus Type 1 Gag-Gag Multimerization in Mammalian Cells{triangledown}

Hua Li,{dagger} Jun Dou, Lingmei Ding, and Paul Spearman*

Departments of Pediatrics and Microbiology and Immunology, Emory University, Atlanta, Georgia 30322

Received 12 June 2007/ Accepted 7 September 2007

The Gag protein of human immunodeficiency virus type 1 directs the virion assembly process. Gag proteins must extensively multimerize during the formation of the spherical immature virion shell. In vitro, virus-like particles can be generated from Gag proteins that lack the N-terminal myristic acid modification or the nucleocapsid (NC) protein. The precise requirements for Gag-Gag multimerization under conditions present in mammalian cells, however, have not been fully elucidated. In this study, a Gag-Gag multimerization assay measuring fluorescence resonance energy transfer was employed to define the Gag domains that are essential for homomultimerization. Three essential components were identified: protein-protein interactions contributed by residues within both the N- and C-terminal domains of capsid (CA), basic residues in NC, and the presence of myristic acid. The requirement of myristic acid for multimerization was reproduced using the heterologous myristoylation sequence from v-src. Only when a leucine zipper dimerization motif was placed in the position of NC was a nonmyristoylated Gag protein able to multimerize. These results support a three-component model for Gag-Gag multimerization that includes membrane interactions mediated by the myristoylated N terminus of Gag, protein-protein interactions between CA domains, and NC-RNA interactions.


* Corresponding author. Mailing address: Pediatric Infectious Diseases, Emory University, 2015 Uppergate Drive, Atlanta, GA 30322. Phone: (404) 727-5642. Fax: (404) 727-9223. E-mail: paul.spearman{at}emory.edu

{triangledown} Published ahead of print on 19 September 2007.

{dagger} Present address: Wistar Institute, Philadelphia, PA 19104.


Journal of Virology, December 2007, p. 12899-12910, Vol. 81, No. 23
0022-538X/07/$08.00+0     doi:10.1128/JVI.01280-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.




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