Mammalian CAP interacts with CAP, CAP2, and actin

Andrew Hubberstey, Gang Yu, Robbie Loewith, Cherelyn Lakusta, Dallan Young

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)459-466
Number of pages8
JournalJournal of Cellular Biochemistry
Volume61
Issue number3
DOIs
StatePublished - 1996

Fingerprint

Yeast
Actins
Yeasts
Cell Extracts
Epitopes
Cells
Peptides
Proteins
Adenylyl Cyclases
Fusion reactions
cytoplasmic antiproteinase
Molecules

Keywords

  • adenylyl cyclase
  • BAT3
  • cytoskeleton
  • RAS
  • signaling
  • yeast

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Mammalian CAP interacts with CAP, CAP2, and actin. / Hubberstey, Andrew; Yu, Gang; Loewith, Robbie; Lakusta, Cherelyn; Young, Dallan.

In: Journal of Cellular Biochemistry, Vol. 61, No. 3, 1996, p. 459-466.

Research output: Contribution to journalArticle

Hubberstey, Andrew ; Yu, Gang ; Loewith, Robbie ; Lakusta, Cherelyn ; Young, Dallan. / Mammalian CAP interacts with CAP, CAP2, and actin. In: Journal of Cellular Biochemistry. 1996 ; Vol. 61, No. 3. pp. 459-466.
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