Mammalian CAP interacts with CAP, CAP2, and actin.
We previously identified human CAP, a homolog of the yeast adenylyl cyclase-associated protein. Previous studies suggest that the N-terminal and C-terminal domains of CAP have distinct functions. We have explored the interactions of human CAP with various proteins. First, by performing yeast two-hybrid screens, we have identified peptides from several proteins that interact with the C-terminal and/or the N-terminal domains of human CAP. These peptides include regions derived from CAP and BAT3, a protein with unknown function. We have further shown that MBP fusions with these peptides can associate in vitro with the N-terminal or C-terminal domains of CAP fused to GST. Our observations indicate that CAP contains regions in both the N-terminal and C-terminal domains that are capable of interacting with each other or with themselves. Furthermore, we found that myc-epitope-tagged CAP coimmunoprecipitates with HA-epitope-tagged CAP from either yeast or mammalian cell extracts. Similar results demonstrate that human CAP can also interact with human CAP2. We also show that human CAP interacts with actin, both by the yeast two-hybrid test and by coimmunoprecipitation of epitope-tagged CAP from yeast or mammalian cell extracts. This interaction requires the C-terminal domain of CAP, but not the N-terminal domain. Thus CAP appears to be capable of interacting in vivo with other CAP molecules, CAP2, and actin. We also show that actin co-immunoprecipitates with HA-CAP2 from mammalian cell extracts.
Hubberstey
A
A
Department of Medical Biochemistry, University of Calgary Health Science Centre, Alberta, Canada.
Yu
G
G
Loewith
R
R
Lakusta
C
C
Young
D
D
D016475:3T3 Cells; D000199:Actins / Q000737:chemistry / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D029721:Drosophila Proteins; D000939:Epitopes / Q000276:immunology; D005656:Fungal Proteins / Q000737:chemistry*; D017405:Herpes Simplex Virus Protein Vmw65 / Q000737:chemistry; D006801:Humans; D051379:Mice; D008840:Microfilament Proteins; D018832:Molecular Chaperones; D008969:Molecular Sequence Data; D009687:Nuclear Proteins; D010957:Plasmids; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry; D012568:Schizosaccharomyces / Q000737:chemistry
D000199:Actins; C092009:BAG6 protein, human; C484266:Bag6 protein, mouse; C489549:CAP1 protein, human; C410502:Capt protein, Drosophila; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D029721:Drosophila Proteins; D000939:Epitopes; D005656:Fungal Proteins; D017405:Herpes Simplex Virus Protein Vmw65; D008840:Microfilament Proteins; D018832:Molecular Chaperones; D009687:Nuclear Proteins; D011506:Proteins