A5080566581- Microtubule and mitosis dynamics
- Nuclear Structure and Function
- RNA Research and Splicing
- Cellular Mechanics and Interactions
- Endoplasmic Reticulum Stress and Disease
- Cellular transport and secretion
- Advanced Fluorescence Microscopy Techniques
- Respiratory Support and Mechanisms
- Genetic and Kidney Cyst Diseases
- Photosynthetic Processes and Mechanisms
- Neonatal Respiratory Health Research
- COVID-19 Clinical Research Studies
- RNA regulation and disease
- Genomics and Chromatin Dynamics
- Protist diversity and phylogeny
- Viral Infectious Diseases and Gene Expression in Insects
- Skin and Cellular Biology Research
- SARS-CoV-2 and COVID-19 Research
- Sports, Gender, and Society
- Advanced Electron Microscopy Techniques and Applications
- CRISPR and Genetic Engineering
- Music History and Culture
- Plant nutrient uptake and metabolism
- Archaeology and ancient environmental studies
- Mitochondrial Function and Pathology
Nikon (United States)
2020-2024
University of California, San Diego
2020-2024
Imaging Center
2020-2024
Altos Labs
2024
La Jolla Alcohol Research
2023
University of Dundee
2011-2018
MRC Protein Phosphorylation and Ubiquitylation Unit
2018
National Institutes of Health
2018
Marine Biological Laboratory
2009-2015
Wellcome Trust
2015
Nucleoporin 98 (Nup98), a glycine-leucine-phenylalanine-glycine (GLFG) amino acid repeat-containing nucleoporin, plays critical part in nuclear trafficking. Injection of antibodies to Nup98 into the nucleus blocks export most RNAs. contains binding sites for several transport factors; however, mechanism by which this nucleoporin functions has remained unclear. Multiple subcellular localizations have been suggested Nup98. Here we show that is indeed found both at pore complex and within...
The eukaryotic spindle assembly checkpoint (SAC) monitors microtubule attachment to kinetochores and prevents anaphase onset until all are aligned on the metaphase plate. In higher eukaryotes, cytoplasmic dynein is involved in silencing SAC by removing proteins Mad2 Rod–Zw10–Zwilch complex (RZZ) from (Howell, B.J., B.F. McEwen, J.C. Canman, D.B. Hoffman, E.M. Farrar, C.L. Rieder, E.D. Salmon. 2001. J. Cell Biol. 155:1159–1172; Wojcik, E., R. Basto, M. Serr, F. Scaerou, Karess, T. Hays. Nat....
The vertebrate nuclear pore is an enormous structure that spans the double membrane of envelope. In yeast, most nucleoporins are found symmetrically on both and cytoplasmic sides structure. However, in vertebrates have been localized exclusively to one side pore. Herein, we show, by immunofluorescence immunoelectron microscopy, Nup98 complex. Additionally, find pore-targeting domain interacts directly with nucleoporin Nup88, a component Nup214, Nup62 subcomplex. was previously described...
BACKGROUND. Multisystem inflammatory syndrome in children (MIS-C) is a rare but potentially severe illness that follows exposure to acute respiratory coronavirus 2 (SARS-CoV-2). Kawasaki disease (KD) shares several clinical features with MIS-C, which prompted the use of intravenous immunoglobulin (IVIG), mainstay therapy for KD. Both diseases share robust activation innate immune system, including IL-1 signaling pathway, and blockade has been used treatment both MIS-C The mechanism action...
The interactions between antibodies, SARS-CoV-2 and immune cells contribute to the pathogenesis of COVID-19 protective immunity. To understand differences antibody responses in mild versus severe cases COVID-19, we analyzed B cell patients 1.5 months post infection. Severe, not mild, infection correlated with high titers IgG against Spike receptor binding domain (RBD) that were capable ACE2:RBD inhibition. (BCR) sequencing revealed VH3-53 was enriched during Of 22 antibodies cloned from two...
Despite the apparent overall structural stability of nuclear pore complex during interphase, at least two nucleoporins have been shown to move dynamically on and off pore. It is not yet certain what contribution nucleoporin mobility makes process transport or how such regulated. Previously, we showed that Nup98 interacts with NPC as well bodies within nucleus in a transcription-dependent manner. We extended our studies dynamics include Nup153, another mobile implicated RNA export. In both...
ABSTRACT Our previous studies of PAWS1 (protein associated with SMAD1; also known as FAM83G) have suggested that this molecule has roles beyond BMP signalling. To investigate these roles, we used CRISPR/Cas9 to generate PAWS1-knockout U2OS osteosarcoma cells. Here, show plays a role in the regulation cytoskeletal machinery, including actin and focal adhesion dynamics, cell migration. Confocal microscopy live imaging cells indicate is involved dynamics organization. Loss causes severe defects...
The formation of the mitotic spindle is a complex process that requires massive cellular reorganization. Regulation by kinases controls this entire process. One these controllers Aurora A kinase, which itself highly regulated. In study, we show nuclear pore protein ALADIN novel spatial regulator A. Without ALADIN, spreads from centrosomes onto microtubules, affects distribution subset microtubule regulators and slows assembly chromosome alignment. interacts with inactive recruited to pole...
The endoplasmic reticulum (ER) is composed of sheets and tubules. Here we report that the COPII coat subunit, SEC24C, works with long form tubular ER-phagy receptor, RTN3, to target dominant-interfering mutant proinsulin Akita puncta lysosomes. When delivery lysosomes was disrupted, large accumulated in ER. Unexpectedly, photobleach analysis indicated behaved as condensates not aggregates, previously suggested. enlarged when either RTN3 or SEC24C were depleted, ER proliferated by knocking...
Neutrophil extracellular traps (NETs) not only counteract bacterial and fungal pathogens but can also promote thrombosis, autoimmunity, sterile inflammation. The presence of citrullinated histones, generated by the peptidylarginine deiminase 4 (PAD4), is synonymous with NETosis considered independent apoptosis. Mitochondrial- death receptor–mediated apoptosis gasdermin E (GSDME)–dependent calcium mobilization membrane permeabilization leading to histone H3 citrullination (H3Cit), nuclear DNA...
The prevalence of "long COVID" is just one the conundrums highlighting how little we know about lung's response to viral infection, particularly syndromecoronavirus-2 (SARS-CoV-2), for which lung point entry. We used an in vitro human system enable a prospective, unbiased, sequential single-cell level analysis pulmonary cell responses infection by multiple SARS-CoV-2 strains. Starting with induced pluripotent stem cells and emulating organogenesis, generated infected three-dimensional,...
Signals from the mitotic spindle during anaphase specify location of actomyosin contractile ring cytokinesis, but detailed mechanism remains unresolved. Here, we have imaged dynamics green fluorescent protein-tagged myosin filaments, microtubules, and Kinesin-6 (which carries activators Rho guanosine triphosphatase) at cell cortex using total internal reflection fluorescence microscopy in flattened Drosophila S2 cells. At onset, relocalizes to microtubule plus ends that grow toward cortex,...
Gle1 is required for mRNA export in yeast and human cells. Here, we report that two (hGle1) isoforms are expressed HeLa cells (hGle1A B). The encoded proteins identical except their COOH-terminal regions. hGle1A ends with a unique four–amino acid segment, whereas hGle1B has 43–amino span. Only hGle1B, the more abundant isoform, localizes to nuclear envelope (NE) pore complex. To test whether hGle1 dynamic shuttling transport factor, microinjected recombinant conducted photobleaching studies...
We identify the prolyl-tRNA synthetase (PRS) inhibitor halofuginone 1 , a compound in clinical trials for anti-fibrotic and anti-inflammatory applications 2 as potent of SARS-CoV-2 infection replication. The interaction spike protein with cell surface heparan sulfate (HS) promotes viral entry 3 . find that reduces HS biosynthesis, thereby reducing binding, pseudotyped virus, authentic infection. Halofuginone also potently suppresses replication post-entry is 1,000-fold more than Remdesivir 4...
ABSTRACT A key event in deflagellation or deciliation is the severing of nine outer-doublet axonemal microtubules at a specific site flagellar transition zone. Previous genetic analysis revealed three genes that are essential for Chlamydomonas. We have now identified first these products, Fa1p, protein required Ca2+-dependent, microtubule severing. Genetic mapping and availability tagged allele allowed us to physically map gene centromere-proximal domain mating-type locus. clones...
PHD1 (also known as EGLN2) belongs to a family of prolyl hydroxylases (PHDs) that are involved in the control cellular response hypoxia. is also able regulate mitotic progression through regulation crucial centrosomal protein Cep192, establishing link between oxygen-sensing and cell cycle machinery. Here, we demonstrate phosphorylated by CDK2, CDK4 CDK6 at S130. This phosphorylation fluctuates with can be induced oncogenic activation. Functionally, leads increased induction hypoxia-inducible...
Dynein is the sole processive minus-end directed microtubule motor found in animals. It has roles cell division, membrane trafficking, and migration. Together with dynactin, dynein regulates centrosomal orientation to establish maintain polarity, controls focal adhesion turnover anchors microtubules at leading edge. In higher eukaryotes, dynein/dynactin requires additional components such as Bicaudal D form an active complex for regulating its cellular localization. Spindly a protein that...
Asymmetric cell divisions depend on the precise placement of spindle apparatus. In mammalian oocytes, spindles assemble close to cell's center, but chromosome segregation takes place at periphery where half chromosomes are expelled into small, nondeveloping polar bodies anaphase. By dividing so asymmetrically, most cytoplasmic content within oocyte is preserved, which critical for successful fertilization and early development. Recently we determined that nucleoporin ALADIN participates in...
Spindly was originally identified as a specific regulator of Dynein activity at the kinetochore. In early prometaphase, recruits Dynein/Dynactin complex, promoting establishment stable kinetochore-microtubule interactions and progression into anaphase. While details function in mitosis have been worked out cultured human cells C. elegans zygote, within context an organism has not yet addressed. Here, we present loss- gain-of-function studies using transgenic RNAi Drosophila. Knock-down...
Abstract Ubiquitylation is a protein modification implicated in several cellular processes. This process reversible by the action of deubiquinating enzymes (DUBs). USP45 ubiquitin specific protease about which little known, aside from roles DNA damage repair and differentiation vertebrate retina. Here, using mass spectrometry we have identified Spindly as new target USP45. Our data show that are part same complex their interaction specifically depends on catalytic activity In addition,...