Biochemical characterization of a family of proteins that stabilizes a plant Ran protein in its GTP-bound conformation

Abstract

Ran-binding proteins (RanBP) are a group of proteins that bind to Ran (Ras-related nuclear small G-protein) and thus control the GTP/GDP-bound states of the Ran and couple the Ran GTPase cycle to cellular processes. In an effort to identify potential downstream effectors for PsRan1-dependent cellular processes, we detected a group of pea Ran (PsRan1)-binding proteins and characterized their biochemical activities. A Ran overlay assay using [ 32P-GTP]-labeled PsRan1 revealed three PsRan1-binding proteins (33, 45, and 85 kDa in size) from total protein extracts of dark-grown pea plumules. These proteins bound preferentially to the Ran-GTP over Ran-GDP conformation and subsequently stabilized its GTP-bound status. We propose that they are a family of proteins that maintain the Ran protein in the active conformation and are potential downstream mediators for PsRan1-dependent cellular processes. Our report provides the basis for characterizing and dissecting Ran downstream targets and Ran-mediated events, and it thus facilitates our understanding about the roles played by Ran/RanBP signaling pathways during plant growth and development.