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(Received for publication, April 25, 1997, and in revised form, May 23, 1997)
From the Tyrosine kinases of the Src family are regulated
via their Src homology 2 (SH2) and SH3 domains. The Nef protein of
human immunodeficiency virus-1 (HIV-1) has previously been shown to bind with high affinity and specificity in vitro to the SH3
domain of Hck, a Src family member expressed primarily in myeloid
cells. However, the effect of Nef on Hck activity in living cells is unknown. Here we show that Rat-2 fibroblasts co-expressing Hck and Nef
rapidly developed transformed foci, whereas control cells expressing
either protein alone did not. Nef formed a stable complex with Hck and
stimulated its tyrosine kinase activity in vivo. Mutagenesis of the Nef proline-rich motif essential for SH3 binding completely blocked complex formation, kinase activation, and
transformation, indicating that the Nef SH3-binding function is
required for its effects on Hck. These results provide direct evidence
that SH3 engagement is sufficient to activate a Src family kinase
in vivo and suggest that Hck may be activated by Nef in
HIV-infected macrophages.
Volume 272, Number 29,
Issue of July 18, 1997
pp. 17899-17902
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
COMMUNICATION:
,
Eppley Institute for Research in Cancer,
University of Nebraska Medical Center, Omaha, Nebraska 68198-6805 and
the § Program in Molecular Medicine, University of
Massachusetts Medical Center, Worcester, Massachusetts 01605
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G. B. Cohen, V. S. Rangan, B. K. Chen, S. Smith, and D. Baltimore The Human Thioesterase II Protein Binds to a Site on HIV-1 Nef Critical for CD4 Down-regulation J. Biol. Chem., July 21, 2000; 275(30): 23097 - 23105. [Abstract] [Full Text] [PDF]
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S. D. Briggs, B. Scholtz, J.-M. Jacque, S. Swingler, M. Stevenson, and T. E. Smithgall HIV-1 Nef Promotes Survival of Myeloid Cells by a Stat3-dependent Pathway J. Biol. Chem., June 29, 2001; 276(27): 25605 - 25611. [Abstract] [Full Text] [PDF]
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A. Manninen, G. Herma Renkema, and K. Saksela Synergistic Activation of NFAT by HIV-1 Nef and the Ras/MAPK Pathway J. Biol. Chem., May 26, 2000; 275(22): 16513 - 16517. [Abstract] [Full Text] [PDF]
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