A5038795878- Cell Image Analysis Techniques
- Research Data Management Practices
- Scientific Computing and Data Management
- Nuclear Structure and Function
- RNA Research and Splicing
- Single-cell and spatial transcriptomics
- Genomics and Chromatin Dynamics
- HIV Research and Treatment
- Genetics, Bioinformatics, and Biomedical Research
- Mass Spectrometry Techniques and Applications
- Advanced Fluorescence Microscopy Techniques
- Radiomics and Machine Learning in Medical Imaging
- AI in cancer detection
- Advanced Proteomics Techniques and Applications
- Molecular Biology Techniques and Applications
- Biomedical and Engineering Education
- RNA regulation and disease
- Metabolomics and Mass Spectrometry Studies
- Signaling Pathways in Disease
- Immune Cell Function and Interaction
- interferon and immune responses
- Cytomegalovirus and herpesvirus research
- Neonatal Health and Biochemistry
- Immunotherapy and Immune Responses
- Biochemical and Molecular Research
University of Massachusetts Chan Medical School
2014-2024
European Molecular Biology Organization
2023
University of Massachusetts Amherst
2021
Institute of Molecular Medicine
2013
Massachusetts Academy of Math and Science
2013
University of Geneva
2010-2011
Rockefeller University
1995-2005
Howard Hughes Medical Institute
1995-1999
University of Alabama at Birmingham
1992
While much has been learned in recent years about the movement of soluble transport factors across nuclear pore complex (NPC), comparatively little is known intranuclear trafficking. We isolated previously identified Saccharomyces protein Mlp1p (myosin-like protein) by an assay designed to find envelope (NE) associated proteins that are not nucleoporins. localized both and a closely related we termed Mlp2p filamentous structures stretching from nucleoplasmic face NE into nucleoplasm, similar...
As tryptic peptides and metabolites are not equally distributed along the mass range, probability of cross fragment ion interference is higher in certain windows when fixed Q1 SWATH applied. We evaluated benefits utilizing variable with regards to selectivity improvement. Variable based on equalizing distribution either precursor population (PIP) or total current (TIC) within each window were generated by an in-house software, swathTUNER. These two strategies outperformed, respect...
The basket of the nuclear pore complex (NPC) is generally depicted as a discrete structure eight protein filaments that protrude into nucleoplasm and converge in ring distal to NPC. We show yeast proteins Mlp1p Mlp2p are necessary components they also embed NPC within dynamic network, whose extended interactome includes spindle organizer, silencing factors, proteasome, key messenger ribonucleoproteins (mRNPs). Ultrastructural observations indicate reduces chromatin crowding around central...
We have identified a new yeast nucleoporin of 159 kDa that we term NUP159. Immunofluorescence microscopy with monospecific monoclonal antibody against NUP159 gave the punctate nuclear rim staining characteristic nucleoporins. Immunogold electron isolated NEs yielded decoration only cytoplasmic side pore complex. The gene encoding is essential, and, like some other nucleoporins, contains coiled-coil domain as well repeated motifs. Five segments NUP159, covering its entire length, were...
New World monkeys of the genus Aotus synthesize a fusion protein (AoT5Cyp) containing tripartite motif-containing 5 (TRIM5) and cyclophilin A (CypA) that potently blocks HIV-1 infection. We attempted to generate human inhibitor modeled after AoT5Cyp, by fusing CypA TRIM5 (hT5Cyp). Of 13 constructs, 3 showed substantial HIV-1-inhibitory activity when expressed in cell lines. This required capsid binding correlated with linkage TRIM5a capsid-specificity determinant ability form cytoplasmic...
Abstract Background Uncoating of the HIV-1 core plays a critical role during early post-fusion stages infection but is poorly understood. Microscopy-based assays are unable to easily distinguish between intact and partially uncoated viral cores. Results In this study, we used 5-ethynyl uridine (EU) label viral-associated RNA HIV production. At time points after with EU-labeled virions, was stained an EU-specific dye detected by confocal microscopy together proteins. We observed that...
The major capsid (CA) protein of retroviruses possesses a stretch 20 amino acids, called the homology region (MHR), which is evolutionarily conserved and invariant in location within primary sequence protein. function this was investigated by examining effect random single-amino-acid substitutions central 13 positions MHR on life cycle Mason-Pfizer monkey virus (M-PMV), an immunosuppressive D-type retrovirus. When these mutants were subcloned into M-PMV proviral vector expressed COS cells,...
A modern day light microscope has evolved from a tool devoted to making primarily empirical observations what is now sophisticated , quantitative device that an integral part of both physical and life science research. Nowadays, microscopes are found in nearly every experimental laboratory. However, despite their prevalent use capturing quantifying scientific phenomena, neither thorough understanding the principles underlying imaging techniques nor appropriate knowledge how calibrate,...
The two yeast proteins Mlp1p and Mlp2p (homologues of the vertebrate protein Tpr) are filamentous attached to nuclear face pore complexes. Here we perform a proteomic analysis, which reveals that Mlps have strikingly different interacting partners, testifying their roles within cell. We find binds directly Spc110p, Spc42p, Spc29p, three core components spindle pole body (SPB), envelope–associated organizer. further show SPB function is compromised in mlp2 mutants. Cells lacking form...
We have developed a large scale enrichment procedure to prepare yeast nuclear envelopes (NEs). These NEs can be stripped of peripheral proteins produce heparin-extracted NE (H-NE) fraction highly enriched in integral membrane proteins. Extraction H-NEs with detergents revealed previously uncharacterized ring structures associated the that apparently stabilize grommets pore complexes (NPCs). The high yields obtained throughout fractionation allowed balance-sheet tabulation subcellular...
Abstract For quality, interpretation, reproducibility and sharing value, microscopy images should be accompanied by detailed descriptions of the conditions that were used to produce them. Micro-Meta App is an intuitive, highly interoperable, open-source software tool was developed in context 4D Nucleome (4DN) consortium designed facilitate extraction collection relevant metadata as specified recent 4DN-BINA-OME tiered-system Microscopy Metadata specifications. In addition substantially...
Open and practical exchange, dissemination, reuse of specimens data have become a fundamental requirement for life sciences research. The quality the obtained thus findings knowledge derived is significantly influenced by samples, experimental methods, analysis. Therefore, comprehensive precise documentation pre-analytical conditions, analytical procedures, processing are essential to be able assess validity research results. With increasing importance reuse, sharing procedures required that...
We sublocalized the yeast nucleoporin Nup82 to cytoplasmic side of nuclear pore complex (NPC) by immunoelectron microscopy. Moreover, in vitro binding assays we showed that interacts with C-terminal region Nup159, a previously was also localized NPC. Hence, two nucleoporins, and form cytoplasmically oriented subcomplex is likely be part fibers emanating from ring Overexpression Rss1/Gle1, putative and/or mRNA transport factor, shown partially rescue depletion Nup159. show here overexpression...