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Mol. Cell. Biol., 02 1996, 548-556, Vol 16, No. 2
NL Freeman, T Lila, KA Mintzer, Z Chen, AJ Pahk, R Ren, DG Drubin and J Field
Saccharomyces cerevisiae cyclase-associated protein (CAP or Srv2p) is
multifunctional. The N-terminal third of CAP binds to adenylyl cyclase and
has been implicated in adenylyl cyclase activation in vivo. The widely
conserved C-terminal domain of CAP binds to monomeric actin and serves an
important cytoskeletal regulatory function in vivo. In addition, all CAP
homologs contain a centrally located proline-rich region which has no
previously identified function. Recently, SH3 (Src homology 3) domains were
shown to bind to proline-rich regions of proteins. Here we report that the
proline-rich region of CAP is recognized by the SH3 domains of several
proteins, including the yeast actin-associated protein Abp1p.
Immunolocalization experiments demonstrate that CAP colocalizes with
cortical actin-containing structures in vivo and that a region of CAP
containing the SH3 domain binding site is required for this localization.
We also demonstrate that the SH3 domain of yeast Abp1p and that of the
yeast RAS protein guanine nucleotide exchange factor Cdc25p complex with
adenylyl cyclase in vitro. Interestingly, the binding of the Cdc25p SH3
domain is not mediated by CAP and therefore may involve direct binding to
adenylyl cyclase or to an unidentified protein which complexes with
adenylyl cyclase. We also found that CAP homologous from
Schizosaccharomyces pombe and humans bind SH3 domains. The human protein
binds most strongly to the SH3 domain from the abl proto-oncogene. These
observations identify CAP as an SH3 domain-binding protein and suggest that
CAP mediates interactions between SH3 domain proteins and monomeric actin.
Copyright © 1996, American Society for Microbiology
A conserved proline-rich region of the Saccharomyces cerevisiae cyclase- associated protein binds SH3 domains and modulates cytoskeletal localization
Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia 19104, USA.
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