A5035857534- Cellular transport and secretion
- Fungal and yeast genetics research
- Endoplasmic Reticulum Stress and Disease
- Microtubule and mitosis dynamics
- Autophagy in Disease and Therapy
- RNA regulation and disease
- Cellular Mechanics and Interactions
- Retinal Development and Disorders
- Pancreatic function and diabetes
- Viral Infectious Diseases and Gene Expression in Insects
- Genetics, Aging, and Longevity in Model Organisms
- Biotin and Related Studies
- CRISPR and Genetic Engineering
- Gene Regulatory Network Analysis
- Mitochondrial Function and Pathology
- Plant Reproductive Biology
- Pluripotent Stem Cells Research
- Photosynthetic Processes and Mechanisms
- Lipid metabolism and biosynthesis
- Lipid Membrane Structure and Behavior
- RNA Interference and Gene Delivery
- Adenosine and Purinergic Signaling
- Ubiquitin and proteasome pathways
- Cell Adhesion Molecules Research
- Neurobiology and Insect Physiology Research
University of Michigan
2014-2024
Michigan United
2018
University of North Carolina at Chapel Hill
2011-2014
University of California, Los Angeles
2002-2007
University of California, Berkeley
2001-2006
Cellular energy influences all aspects of cellular function. Although cells can adapt to a gradual reduction in energy, acute depletion poses unique challenge. Because hampers the transport new sources into cell, cell must use endogenous substrates replenish after depletion. In yeast Saccharomyces cerevisiae, glucose starvation causes an intracellular that recovers during continued starvation. However, how replenishes early phase is unknown. this study, we investigated role pathways deliver...
Transcription errors occur in all living cells; however, it is unknown how these affect cellular health. To answer this question, we monitor yeast cells that are genetically engineered to display error-prone transcription. We discover suffer from a profound loss proteostasis, which sensitizes them the expression of genes associated with protein-folding diseases humans; thus, transcription represent new molecular mechanism by can acquire disease phenotypes. further find error rate increases...
With the advent of complete genome sequences, large-scale functional analyses are generating new excitement in biology and medicine. To facilitate genomewide analyses, we developed a high-density cell array with quantitative automated readout fitness. Able to print at >×10 higher density on standard microtiter plate area than currently possible, our allows single-plate screening set Saccharomyces cerevisiae gene-deletion library significantly reduces amount small molecules other materials...
Lipid droplet (LD) utilization is an important cellular activity that regulates energy balance and release of lipid second messengers. Because fatty acids exhibit both beneficial toxic properties, their from LDs must be controlled. Here we demonstrate yeast Sfh3, unusual Sec14-like phosphatidylinositol transfer protein, LD-associated protein inhibits mobilization these particles. We further document a complex biochemical diversification during sporulation in which Sfh3 select other LD...
Eisosomes are membrane furrows at the cell surface of yeast that have been shown to function in two seemingly distinct pathways, stress response and regulation nutrient transporters. We found many conditions affect both these pathways by changing plasma tension thus morphology composition eisosomes. For example, alkaline causes swelling an endocytic response, which together increase tension, thereby flattening The flattened eisosomes release transporters, aids their down-regulation. In...
Actin and microtubule dynamics must be precisely coordinated during cell migration, mitosis, morphogenesis--much of this coordination is mediated by proteins that physically bridge the two cytoskeletal networks. We have investigated regulation Drosophila actin-microtubule cross-linker Short stop (Shot), a member spectraplakin family. Our data suggest Shot's cross-linking activity regulated an intramolecular inhibitory mechanism. In its inactive conformation, Shot adopts "closed" conformation...
Abstract Chemical sensitivity, growth inhibition in response to a chemical, is powerful phenotype that can reveal insight into diverse cellular processes. sensitivity assays are used nearly every model system, however the yeast Saccharomyces cerevisiae provides particularly platform for discovery and mechanistic from chemical assays. Here we describe simple inexpensive approach determine quantitatively form of half maximal inhibitory concentration (IC 50 ) using common laboratory equipment....
Critical early steps in human embryonic development include polarization of the inner cell mass, followed by formation an expanded lumen that will become epiblast cavity. Recently described three-dimensional (3D) pluripotent stem cell-derived cyst (hPSC-cyst) structures can replicate these processes. To gain mechanistic insights into poorly understood machinery involved cavity formation, we interrogated proteomes apical and basolateral membrane territories 3D hPSC-cysts. APEX2-based...
Clathrin adaptors are key factors in clathrin-coated vesicle formation, coupling clathrin to cargo and/or the lipid bilayer. A physically interacting network of three classes participate clathrin-mediated traffic between trans-Golgi (TGN) and endosomes: AP-1, Gga proteins, epsin-like proteins. Here we investigate functional relationships within this through transport assays protein localization analysis living yeast cells. We observed that Ent3p preferentially localized with Gga2p, whereas...
Abstract Epitope tags are commonly used for various purposes in research labs. The DYKDDDDK-peptide epitope, trademarked as the FLAG is a epitope tag. It often monitoring protein levels and affinity chromatography. Multiple DYKDDDDK-binding antibodies available; however, mouse monoclonal anti-FLAG M2 widely due to its commercial availability several formats. Many laboratory Saccharomyces cerevisiae strains, including BY4741 strain that was multiple systematic deletion tagging libraries, have...
Control of actin assembly nucleated by the Arp2/3 complex plays a crucial role during budding yeast endocytosis. The Eps15-related activator, Pan1p, is essential for endocytic internalization and proper organization. Pan1p activity negatively regulated Prk1 kinase phosphorylation after internalization. Phosphorylated probably then dephosphorylated in cytosol. recruited to sites approximately 25 s before initiation polymerization, suggesting that its activation kept inactive early stages...
Glucose is a master regulator of cell behavior in the yeast Saccharomyces cerevisiae. It acts as both metabolic substrate and potent intracellular signaling cascades. starvation induces transient delocalization then partial relocalization clathrin adaptors at trans-Golgi network endosomes. Although these localization responses are known to depend on protein kinase A (PKA) pathway, molecular mechanism this regulation unknown. Here we demonstrate that PKA AMP-regulated regulate adaptor through...
Glucose is a rich source of energy and the raw material for biomass increase. Many eukaryotic cells remodel their physiology in presence absence glucose. The yeast Saccharomyces cerevisiae undergoes changes transcription, translation, metabolism, cell polarity response to glucose availability. Upon starvation, translation initiation are immediately inhibited, then gradually recover. In this paper, we provide evidence that, as traffic at trans-Golgi network (TGN) endosomes regulated by via an...
Small molecule inhibitors provide powerful tools to characterize highly dynamic and complex eukaryotic cell pathways such as those mediating membrane traffic. However, a lack of easy generalizable assays has constrained identification novel despite availability diverse chemical libraries. Here, we report facile growth-based strategy in yeast screen for pathway-specific inhibitors. The approach uses well characterized synthetic genetic growth defects guide design cells genetically sensitized...
Background Information In the yeast Saccharomyces cerevisiae, acute glucose starvation induces rapid endocytosis followed by vacuolar degradation of many plasma membrane proteins. This process is essential for cell viability, but regulatory mechanisms that control it remain poorly understood. Under normal growth conditions, a major decision endocytic cargo occurs at trans‐Golgi network (TGN) where proteins can recycle back to or be recognized TGN‐localised clathrin adaptors direct them...
Protein sorting at the trans-Golgi network (TGN) usually requires assistance of cargo adaptors. However, it remains to be examined how same complex can mediate both export and retention different proteins or complexes interact among themselves. In Saccharomyces cerevisiae, exomer is involved in polarized transport some from TGN plasma membrane (PM). Intriguingly, its cargos also show a sort functional relationship with clathrin adaptors that still unsolved. Here, using wide range techniques,...
Clathrin is a ubiquitous protein that mediates membrane traffic at many locations. To function, clathrin requires adaptors link it to transmembrane cargo. In addition this cargo selection also play mechanistic roles in the formation of transport carrier. However, full spectrum these poorly understood. Here we report Ent5, an endosomal adaptor Saccharomyces cerevisiae, regulates behavior coats after recruitment clathrin. We show loss Ent5 disrupts clathrin-dependent and prolongs lifespan...
Membrane traffic is an essential process that allows protein and lipid exchange between the endocytic, lysosomal, secretory compartments. Clathrin-mediated trans-Golgi network endosomes mediates responses to environment through sorting of biosynthetic endocytic cargo. Traffic this pathway initiated by controlled assembly a clathrin-adaptor coat on cytosolic surface originating organelle. In process, clathrin recruited different adaptor proteins act as bridge transmembrane cargo be...