Wsc1 and Mid2 are cell surface sensors for cell wall integrity signaling that act through Rom2, a guanine nucleotide exchange factor for Rho1.
Wsc1 and Mid2 are highly O-glycosylated cell surface proteins that reside in the plasma membrane of Saccharomyces cerevisiae. They have been proposed to function as mechanosensors of cell wall stress induced by wall remodeling during vegetative growth and pheromone-induced morphogenesis. These proteins are required for activation of the cell wall ... integrity signaling pathway that consists of the small G-protein Rho1, protein kinase C (Pkc1), and a mitogen-activated protein kinase cascade. We show here by two-hybrid experiments that the C-terminal cytoplasmic domains of Wsc1 and Mid2 interact with Rom2, a guanine nucleotide exchange factor (GEF) for Rho1. At least with regard to Wsc1, this interaction is mediated by the Rom2 N-terminal domain. This domain is distinct from the Rho1-interacting domain, suggesting that the GEF can interact simultaneously with a sensor and with Rho1. We also demonstrate that extracts from wsc1 and mid2 mutants are deficient in the ability to catalyze GTP loading of Rho1 in vitro, providing evidence that the function of the sensor-Rom2 interaction is to stimulate nucleotide exchange toward this G-protein. In a related line of investigation, we identified the PMT2 gene in a genetic screen for mutations that confer an additive cell lysis defect with a wsc1 null allele. Pmt2 is a member of a six-protein family in yeast that catalyzes the first step in O mannosylation of target proteins. We demonstrate that Mid2 is not mannosylated in a pmt2 mutant and that this modification is important for signaling by Mid2.
Mesh Terms:
Amino Acid Sequence, Calcium-Binding Proteins, Cell Wall, Enzyme Activation, Guanine Nucleotide Exchange Factors, Intracellular Signaling Peptides and Proteins, Mannosyltransferases, Membrane Glycoproteins, Membrane Proteins, Mutation, Pheromones, Protein Binding, Protein Structure, Tertiary, Protein-Serine-Threonine Kinases, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Signal Transduction, Substrate Specificity, Two-Hybrid System Techniques, rho GTP-Binding Proteins
Amino Acid Sequence, Calcium-Binding Proteins, Cell Wall, Enzyme Activation, Guanine Nucleotide Exchange Factors, Intracellular Signaling Peptides and Proteins, Mannosyltransferases, Membrane Glycoproteins, Membrane Proteins, Mutation, Pheromones, Protein Binding, Protein Structure, Tertiary, Protein-Serine-Threonine Kinases, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Signal Transduction, Substrate Specificity, Two-Hybrid System Techniques, rho GTP-Binding Proteins
Mol. Cell. Biol.
Date: Jan. 01, 2001
PubMed ID: 11113201
View in: Pubmed Google Scholar
Download Curated Data For This Publication
17987
Switch View:
- Interactions 4