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Journal of Virology, March 2001, p. 2154-2160, Vol. 75, No. 5
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.5.2154-2160.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Human Immunodeficiency Virus Type 1 Nef Selectively Associates with a Catalytically Active Subpopulation of p21-Activated Kinase 2 (PAK2) Independently of PAK2 Binding to Nck or beta -PIX

G. Herma Renkema,1 Aki Manninen,1 and Kalle Saksela1,2,*

Institute of Medical Technology, University of Tampere,1 and Department of Clinical Chemistry, Tampere University Hospital,2 Tampere, Finland

Received 27 September 2000/Accepted 28 November 2000

We have recently identified the Nef-associated serine-threonine kinase (NAK) as the p21-activated kinase 2 (PAK2). Here we have taken advantage of the possibility to manipulate the functional properties of NAK by transfecting PAK2 cDNA or its mutant derivatives in order to further characterize the Nef-NAK complex. To exclude the possibility that some Nef variants might interact with PAK1 instead of PAK2, we also examined the identity of NAK complexed with divergent human immunodeficiency virus type 1 HIV-1 Nef proteins. All tested Nef proteins, including SF2, NL4-3, BH10, and HAN-2, associated with PAK2 but not with PAK1. By exchanging different regions between these two PAK proteins, the selective ability of PAK2 to associate with Nef could be mapped to the carboxy-terminal part of its regulatory domain. Binding of PAK2 with the adapter protein Nck or beta -PIX was found to be dispensable for the assembly of the Nef-PAK2 complex, whereas an intact Cdc42-Rac1 interactive binding motif was required. Most importantly, we found that NAK represented a distinct subpopulation of the total cellular PAK2 characterized by a high specific kinase activity. Thus, although only a small fraction of cellular PAK2 could be found in complex with Nef, NAK represented a major part of cellular PAK2 activity.

* Corresponding author. Mailing address: Institute of Medical Technology, University of Tampere, P.O. Box 607, FIN-33101 Tampere, Finland. Phone: 358-3-215 7029. Fax: 358-3-215 8597. E-mail: kalle.saksela{at}uta.fi.

Journal of Virology, March 2001, p. 2154-2160, Vol. 75, No. 5
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.5.2154-2160.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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