Two active states of the Ras-related Bud1/Rsr1 protein bind to different effectors to determine yeast cell polarity.
Cells of budding yeast organize their cytoskeleton in a highly polarized manner during vegetative growth. Selection of a site for polarization requires a group of proteins including a Ras-like GTPase, Bud1, and its regulators. Another group of proteins, which includes a Rho-like GTPase (Cdc42), its guanine nucleotide exchange factor (Cdc24), and Bem1, is necessary for organization of the actin cytoskeleton and for cell polarization. We have proposed previously that the Bud1 protein, through its GTPase cycle, determines the localization of one or more of the cell polarity proteins to the bud site. Herein we demonstrate that Bud1 directly interacts with Cdc24 and Bem1: Bud1 in its GTP-bound form associates preferentially with Cdc24, whereas the GDP-bound form of Bud1 associates with Bem1. We also present subcellular fractionation data for Bud1 that is consistent with the idea that Bud1 can travel between the site for budding on the plasma membrane and the cytosol. We propose that Bud1 can exist in two active states for association with different partners and that the switch from Bud1-GTP to Bud1-GDP provides a regulatory device for ordered assembly of a macromolecular complex at the bud site.
Park
H O
HO
Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94143-0448, USA. park.294@osu.edu
Bi
E
E
Pringle
J R
JR
Herskowitz
I
I
D048868:Adaptor Proteins, Signal Transducing; D002451:Cell Compartmentation; D018797:Cell Cycle Proteins / Q000378:metabolism; D016764:Cell Polarity / Q000502:physiology*; D003593:Cytoplasm / Q000737:chemistry; D005656:Fungal Proteins / Q000378:metabolism*; D020558:GTP Phosphohydrolases / Q000235:genetics / Q000378:metabolism*; D020662:Guanine Nucleotide Exchange Factors; D008566:Membranes / Q000737:chemistry; D008954:Models, Biological; D009154:Mutation; D011485:Protein Binding; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins; D015398:Signal Transduction; D020691:rab GTP-Binding Proteins
D048868:Adaptor Proteins, Signal Transducing; C073594:CDC24 protein, S cerevisiae; D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D020662:Guanine Nucleotide Exchange Factors; D011518:Proto-Oncogene Proteins; D029701:Saccharomyces cerevisiae Proteins; C073129:BEM1 protein, S cerevisiae; D020558:GTP Phosphohydrolases; C084013:RSR1 protein, S cerevisiae; D020691:rab GTP-Binding Proteins