Phosphorylation-dependent association of the Ras-related GTP-binding protein Rem with 14-3-3 proteins

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31 Scopus citations


Rem belongs to a subfamily of Ras-related GTPases that includes Rad, Gem, and Kir. These proteins are unique among the Ras superfamily since their expression is under transcriptional regulation and they contain distinct amino and carboxyl termini. To gain insight into the cellular function of Rem, we have undertaken an expression screen using a mouse embryo cDNA library to identify Rem-interacting proteins and find that Rem interacts with a series of 14-3-3 isoforms (ε, φ, and ζ). Immunoprecipitation studies demonstrate an interaction that is independent of the nucleotide state of Rem. Rem is phosphorylated in vivo, and binding of Rem to 14-3-3ζ is abolished by pretreating Rem with protein phosphatase 1. Thus, the association of Rem and 14-3-3ζ is phosphorylation-dependent. Examination of the interaction between 14-3-3ζ and various Rem deletion mutants mapped a critical binding site to the C-terminus of Rem. Finally, we demonstrate the interaction of Rad but not the newly identified Rem2 protein with 14-3-3 proteins. These results suggest that 14-3-3 may allow the recruitment of distinct proteins that participate in Rem-mediated signal transduction pathways.

Original languageEnglish
Pages (from-to)401-412
Number of pages12
JournalArchives of Biochemistry and Biophysics
Issue number2
StatePublished - Aug 15 1999

Bibliographical note

Funding Information:
The nucleotide sequences for rat Rem2 have been submitted to the Genebank Data Bank with Accession No. AF084464. 1This work was supported by NIH Grant EY11231 and a grant from the American Heart Association, Kentucky Affiliate. 2To whom correspondence should be addressed. Fax: 606-323-1037. 3Abbreviations used: GST, glutathione S-transferase; HMK, cAMP-dependent protein kinase from heart muscle; SH2, Src homology 2 domain; GTPase, guanosine triphosphatase; GTPγS, guanosine 5′-3-O-(thio)triphosphate; PAGE, polyacrylamide gel electrophoresis; DTT, dithiothreitol; HA, influenza hemagglutinin epitope; PCR, polymerase chain reaction; BSA, bovine serum albumin; MAP, mi-

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology


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