Stable complex formation between HIV Rev and the nucleosome assembly protein, NAP1, affects Rev function

• Published in: Virology (New York, N.Y.) N.Y.), 2009-05, Vol.388 (1), p.103-111
• Main Authors: Cochrane, Alan, Murley, Laura Lea, Gao, Mian, Wong, Raymond, Clayton, Kiera, Brufatto, Nicole, Canadien, Veronica, Mamelak, Daniel, Chen, Tricia, Richards, Dawn, Zeghouf, Mahel, Greenblatt, Jack, Burks, Christian, Frappier, Lori
• Format: Article
• Language: English
• Subjects: 60 APPLIED LIFE SCIENCES; AFFINITY; AIDS VIRUS; AMINO ACIDS; ANIMAL CELLS; ARGININE; CARBOXYLIC ACIDS; Cell-Free System; Chromatography, Affinity; HeLa Cells; HIV; Human immunodeficiency virus 1; Humans; Infectious Disease; INTERACTIONS; MICROORGANISMS; NUCLEIC ACIDS; Nucleolar localization; Nucleosome assembly protein; ORGANIC ACIDS; ORGANIC COMPOUNDS; PARASITES; PROLIFERATION; PROTEINS; Proteins - metabolism; PURIFICATION; Rev; rev Gene Products, Human Immunodeficiency Virus - genetics; rev Gene Products, Human Immunodeficiency Virus - metabolism; RNA; TAP-tagging; VIRUSES
• ISSN: 0042-6822; 1096-0341
• DOI: 10.1016/j.virol.2009.03.005

Abstract The Rev protein of HIV-1 is essential for HIV-1 proliferation due to its role in exporting viral RNA from the nucleus. We used a modified version of tandem affinity purification (TAP) tagging to identify proteins interacting with HIV-1 Rev in human cells and discovered a prominent interaction between Rev and nucleosome assembly protein 1 (Nap1). This interaction was also observed by specific retention of Nap1 from human cell lysates on a Rev affinity column. Nap1 was found to bind Rev through the Rev arginine-rich domain and altered the oligomerization state of Rev in vitro . Overexpression of Nap1 stimulated the ability of Rev to export RNA, reduced the nucleolar localization of Rev, and affected Rev nuclear import rates. The results suggest that Nap-1 may influence Rev function by increasing the availability of Rev.