A5002286065- Fungal and yeast genetics research
- Microtubule and mitosis dynamics
- CRISPR and Genetic Engineering
- Ubiquitin and proteasome pathways
- Plant-Microbe Interactions and Immunity
- Gene Regulatory Network Analysis
- Plant Reproductive Biology
- Fungal Biology and Applications
- RNA and protein synthesis mechanisms
- Protein Kinase Regulation and GTPase Signaling
- Cellular transport and secretion
- Genomics and Chromatin Dynamics
- Bacterial Genetics and Biotechnology
- Viral Infectious Diseases and Gene Expression in Insects
- Fermentation and Sensory Analysis
- Polyamine Metabolism and Applications
- Melanoma and MAPK Pathways
- Virus-based gene therapy research
- Heat shock proteins research
- HIV Research and Treatment
- RNA Research and Splicing
- HIV/AIDS drug development and treatment
- thermodynamics and calorimetric analyses
- Molecular Biology Techniques and Applications
- Bioinformatics and Genomic Networks
University of Massachusetts Chan Medical School
2013-2025
University of California, San Francisco
1992-1998
University of Washington
1993-1996
University of California San Francisco Medical Center
1989
University of California, Los Angeles
1986
In the Saccharomyces cerevisiae pheromone response pathway, Gbetagamma complex activates downstream responses by an unknown mechanism involving a MAP kinase cascade, PAK-like Ste20, and Rho family GTPase, Cdc42. Here we show that must remain membrane-associated after release from Galpha to activate pathway. We also stimulates translocation of cascade scaffold protein Ste5 cell surface. This recruitment requires function Gbetagamma-binding domain Ste5, but not kinases Gbetagamma, suggesting...
Heterotrimeric guanosine triphosphate (GTP)–binding proteins (G proteins) determine tissue and cell polarity in a variety of organisms. In yeast, cells orient polarized growth toward the mating partner along pheromone gradient by mechanism that requires Far1p Cdc24p. bound Gβγ interacted with establishment proteins, which organize actin cytoskeleton. Cells containing mutated unable to bind or were defective for orienting their partner. response pheromones, moves from nucleus cytoplasm. Thus,...
Abstract Twelve‐ and sixteen‐residue peptides have been designed to form tetrameric alphahelical bundles. Both are capable of folding into amphiphilic alpha‐helices, with leucyl residues along one face glutamyl lysyl the opposite face. Four such alpha‐helices forming a noncovalently bonded tetramer. Neighboring helices run in antiparallel directions design, so that complex has 222 symmetry. In tetramer, side chains interdigitate center hydrophobic interaction, charged exposed solvent. The...
The Saccharomyces cerevisiae kinase Ste20 is a member of the p21-activated (PAK) family with several functions, including pheromone-responsive signal transduction. While PAKs are usually activated by small G proteins and binds Cdc42, role Cdc42-Ste20 binding has been controversial, largely because lacking its entire Cdc42-binding (CRIB) domain retains activity pheromone response. Here we show that, unlike CRIB deletion, point mutations in that disrupt Cdc42 also signaling. We found...
During conjugation, haploid S. cerevisiae cells find one another by polarizing their growth toward each other along gradients of pheromone (chemotropism). We demonstrate that yeast exhibit a second mating behavior: when receptors are saturated with pheromone, wild-type execute default pathway and select mate at random. These matings less efficient than chemotropic matings, induced the same dose induces shmoo formation, appear to use site near incipient bud for polarization. show SPA2 gene is...
Hsp70 interactions are critical for cellular viability and the response to stress. Previous attempts characterize have been limited by their transient nature inability of current technologies distinguish direct versus bridged interactions. We report novel use cross-linking mass spectrometry (XL-MS) comprehensively Saccharomyces cerevisiae (budding yeast) protein interactome. Using this approach, we gained fundamental new insights into function, including definitive evidence self-association...
We have investigated the mechanisms by which alleles at mouse Fv-1 locus restrict replication of murine leukemia viruses. Inhibition productive infection is closely paralleled reduced accumulation integrated proviral DNA as well levels linear viral in a cytoplasmic fraction. Nevertheless, present nearly normal nuclear fraction, and total amounts are only mildly affected restrictive infections, suggesting block integration to account for DNA. However, integrase (IN)-dependent trimming 3' ends...
The Rho-type GTPase Cdc42 is a central regulator of eukaryotic cell polarity and signal transduction. In budding yeast, regulates mitogen-activated protein (MAP) kinase signaling in part through the PAK-family Ste20. Activation Ste20 requires Cdc42/Rac interactive binding (CRIB) domain, which mediates its recruitment to membrane-associated Cdc42. Here, we identify separate domain that interacts directly with membrane phospholipids critical for function. This short region, termed basic-rich...
CDC42 encodes a highly conserved GTPase of the Rho family that is best known for its role in regulating cell polarity and actin organization. In addition, various studies both yeast mammalian cells have suggested Cdc42p, through interaction with p21-activated kinases (PAKs), plays signaling pathways regulate target gene transcription. However, recent pheromone response pathway prior results temperature-sensitive cdc42 mutants were misleading Cdc42p Cdc42p-PAK are not involved signaling. To...
We present a method for studying multiple retroviral integration events into small DNA target in vivo. Episomal simian virus 40 (SV40) genomes established by infection of CV-1 cells served as targets during subsequent with murine leukemia (MLV). Using PCR-based assay the abundance and distribution events, nonrandom MLV SV40 is detectable early 4 hr reaches maximum level 8 after infection. The but not sites sensitive to stage life cycle at which performed. temperature-sensitive tumor (T)...
Mating pheromones of Saccharomyces cerevisiae control both signal transduction events and changes in cell shape. The G beta gamma complex the pheromone receptor-coupled protein activates pathway, leading to transcriptional induction cycle arrest, but how pheromone-dependent signalling leads shape is unclear. We used a two-hybrid system search for proteins that interact with might be involved changes. identified ankyrin repeat-containing Akr1p show here it interacts free complex. This...
The Saccharomyces cerevisiae PAK (p21-activated kinase) family kinase Ste20 functions in several signal transduction pathways, including pheromone response, filamentous growth, and hyperosmotic resistance. GTPase Cdc42 localizes activates by binding to an autoinhibitory motif within called the CRIB domain. Another factor that with is protein Bem1. Bem1 has two SH3 domains, but target ligands for these domains have not been described. Here we identify evolutionarily conserved site between of...