A5036040264- Fungal and yeast genetics research
- Nuclear Structure and Function
- RNA Research and Splicing
- Cellular transport and secretion
- Endoplasmic Reticulum Stress and Disease
- Genomics and Chromatin Dynamics
- Ubiquitin and proteasome pathways
- Trace Elements in Health
- DNA Repair Mechanisms
- Drug Transport and Resistance Mechanisms
- Polyamine Metabolism and Applications
- Bacterial Genetics and Biotechnology
- Biotin and Related Studies
- Autophagy in Disease and Therapy
- Cystic Fibrosis Research Advances
- RNA and protein synthesis mechanisms
- Retinoids in leukemia and cellular processes
- Prion Diseases and Protein Misfolding
- Microbial Natural Products and Biosynthesis
- Fermentation and Sensory Analysis
- Peptidase Inhibition and Analysis
- Plant biochemistry and biosynthesis
- Lysosomal Storage Disorders Research
- HIV/AIDS drug development and treatment
- RNA regulation and disease
Johns Hopkins University
2011-2023
Johns Hopkins Medicine
2006-2023
Columbia University
2018
University of Washington
2005
University of California, San Francisco
1986-2002
University of California, Davis
2002
Gladstone Institutes
2002
Harvard University
1998
Nagoya University
1998
Princeton University
1996
Zmpste24 is an integral membrane metalloproteinase of the endoplasmic reticulum. Biochemical studies tissues from -deficient mice ( −/− ) have indicated a role for in processing CAAX -type prenylated proteins. Here, we report pathologic consequences deficiency mice. gain weight slowly, appear malnourished, and exhibit progressive hair loss. The most striking phenotype multiple spontaneous bone fractures—akin to those occurring mouse models osteogenesis imperfecta. Cortical trabecular volumes...
Prenylcysteine carboxyl methyltransferase (pcCMT) is the third of three enzymes that posttranslationally modify C-terminal CAA<i>X</i> motifs and thereby target proteins to plasma membrane. Here we report molecular characterization subcellular localization first mammalian (human myeloid) pcCMT. The deduced amino acid sequence pcCMT predicts a multiple membrane-spanning protein with homologies yeast pcCMT, <i>STE14</i> , band 3 anion transporter. human gene complemented <i>ste14</i> mutant....
The folding and assembly of proteins in the endoplasmic reticulum (ER) lumen membrane are monitored by ER quality control. Misfolded or unassembled retained and, if they cannot fold assemble correctly, ultimately undergo ER-associated degradation (ERAD) mediated ubiquitin-proteasome system. Whereas luminal integral ERAD substrates both require proteasome for their degradation, control machinery these two classes likely differs because distinct topologies. Here we establish requirements...
Membrane and secretory proteins fold in the endoplasmic reticulum (ER), misfolded may be retained targeted for ER-associated protein degradation (ERAD). To elucidate mechanism by which an integral membrane ER is degraded, we studied fate of cystic fibrosis transmembrane conductance regulator (CFTR) yeast Saccharomyces cerevisiae. Our data indicate that CFTR resides stabilized strains defective proteasome activity or deleted ubiquitin-conjugating enzymes Ubc6p Ubc7p, thus demonstrating a bona...
The Saccharomyces cerevisiae pheromone a-factor is produced by a cells and interacts with alpha to cause cell cycle arrest other physiological responses associated mating. Two structural genes, MFA1 MFA2, have been previously cloned synthetic probes based on the amino acid sequence (A. Brake, C. Brenner, R. Najarian, P. Laybourn, J. Merryweather, cited in M.-J. Gething [ed.], Protein transport secretion, 1985). We examined function of these genes production mating construction analysis...
Hutchinson-Gilford progeria syndrome (HGPS) is a devastating premature aging disease resulting from mutation in the LMNA gene, which encodes nuclear lamins A and C. Lamin synthesized as precursor (prelamin A) with C-terminal CaaX motif that undergoes farnesylation, endoproteolytic cleavage, carboxylmethylation. Prelamin subsequently internally cleaved by zinc metalloprotease Ste24 (Zmpste24) protease, removes 15 amino acids, including modifications, to yield mature lamin A. HGPS results...
The nuclear lamins form a karyoskeleton providing structural rigidity to the nucleus. One member of lamin family, A, is first synthesized as 74 kDa precursor, prelamin A. After endopeptidase and methylation reactions which occur after farnesylation CAAX-box cysteine, there second endoproteolysis that occurs 15 amino acids upstream from C-terminal farnesylated cysteine residue. Studies with knockout mice have implicated enzyme Zmpste24 (Face-1) suitable candidate perform one or both these...
Protein misfolding is monitored by a variety of cellular "quality control" systems. Endoplasmic reticulum (ER) quality control handles misfolded secretory and membrane proteins well characterized. However, less known about the cytosolic (CytoQC). To study CytoQC, we have employed genetic system in Saccharomyces cerevisiae using transplantable degron, CL1 (1). Attachment to protein Ura3p destabilizes Ura3p, targeting it for rapid proteasomal degradation. We performed comprehensive analysis...
Proteins terminating in the CAAX motif, for example Ras and yeast a-factor mating pheromone, are prenylated, trimmed of their last three amino acids, carboxyl-methylated. The enzymes that mediate these activities, collectively referred to as processing components, have been identified genetically Saccharomyces cerevisiae . Whereas Ram1p/Ram2p prenyltransferase is a cytosolic soluble enzyme, sequence analysis predicts other Rce1p Ste24p proteases Ste14p methyltransferase, contain multiple...
In the yeast Saccharomyces cerevisiae, mutations in either of two unlinked genes, RAM1 or RAM2, abolish farnesyltransferase activity responsible for prenylation Ras proteins and a-factor mating pheromone. Here we report that function RAM2 genes is required membrane localization a-factor. The gene was sequenced can encode a 38-kDa protein. We examined functional interaction by expressing Escherichia coli. Extracts derived from an E. coli strain coexpressed efficiently farnesylated peptide...
Many secreted bioactive signaling molecules, including the yeast mating pheromones a-factor and α-factor, are initially synthesized as precursors requiring multiple intracellular processing enzymes to generate their mature forms. To identify new gene products involved in biogenesis of Saccharomyces cerevisiae, we carried out a screen for MATa-specific, mating-defective mutants. We have identified mutant, ste24, addition previously known sterile During its wild-type strain, precursor...
The zinc metalloprotease ZMPSTE24 plays a critical role in nuclear lamin biology by cleaving the prenylated and carboxylmethylated 15-amino acid tail from C-terminus of prelamin A to yield mature A. defect this proteolytic event, caused mutation gene (LMNA) that eliminates cleavage site, underlies premature aging disease Hutchinson-Gilford Progeria Syndrome (HGPS). Likewise, mutations result decreased enzyme function cause spectrum diseases share certain features aging. Twenty human alleles...
Abstract Cellular protein quality control (PQC) systems selectively target misfolded or otherwise aberrant proteins for degradation by the ubiquitin-proteasome system (UPS). How cells discern abnormal from normal remains incompletely understood, but involves in part recognition between ubiquitin E3 ligases and signals (degrons) that are exposed proteins. PQC is compartmentalized cell, a great deal has been learned recent years about ER-associated (ERAD) nuclear control. In contrast,...
Maturation of the Saccharomyces cerevisiae a-factor precursor involves COOH-terminal CAAX processing (prenylation, AAX tripeptide proteolysis, and carboxyl methylation) followed by cleavage an NH2-terminal extension (two sequential proteolytic steps). The aim this study is to clarify precise role Ste24p, a membrane-spanning zinc metalloprotease, in precursor. We demonstrated previously that Ste24p necessary for first step analysis radiolabeled intermediates vivo (Fujimura-Kamada, K., F.J....