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.
-PIX
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
-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.
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