A5009166035- Microtubule and mitosis dynamics
- Cellular transport and secretion
- Protist diversity and phylogeny
- Photosynthetic Processes and Mechanisms
- Epigenetics and DNA Methylation
- Cellular Mechanics and Interactions
- Ubiquitin and proteasome pathways
- Nuclear Structure and Function
- Genetic and Kidney Cyst Diseases
- RNA modifications and cancer
- Mitochondrial Function and Pathology
- Cardiomyopathy and Myosin Studies
- Genomics and Chromatin Dynamics
- Muscle Physiology and Disorders
- Genetics, Aging, and Longevity in Model Organisms
- Cancer-related Molecular Pathways
- Glioma Diagnosis and Treatment
- MicroRNA in disease regulation
- RNA and protein synthesis mechanisms
- Cell Adhesion Molecules Research
- Lysosomal Storage Disorders Research
- Congenital heart defects research
- 14-3-3 protein interactions
- Glycosylation and Glycoproteins Research
- Fungal and yeast genetics research
University of Notre Dame
2011-2024
University of Manchester
2014
University of Massachusetts Chan Medical School
1999-2000
Foundation for Biomedical Research
1995-1997
Washington University in St. Louis
1993-1994
University of Cambridge
1993
Centre National de la Recherche Scientifique
1993
Wellcome Sanger Institute
1993
Universidade de São Paulo
1993
Yale University
1993
Dynactin is a multi-subunit complex which has been implicated in cytoplasmic dynein function, though its mechanism of action unknown. In this study, we have characterized the 50-kD subunit dynactin, and analyzed effects overexpression on mitosis living cells. Rat human cDNA clones revealed p50 to be novel highly conserved, containing three predicted coiled-coil domains. Immunofluorescence staining dynactin components cultured vertebrate cells showed that both complexes are recruited...
Cytoplasmic dynein is a retrograde microtubule motor thought to participate in organelle transport and some aspects of minus end-directed chromosome movement. The mechanism binding organelles kinetochores unknown. Based on homology with the Chlamydomonas flagellar outer arm intermediate chains (ICs), we proposed role for cytoplasmic ICs linking protein kinetochores. In this study two different IC isoforms were used blot overlay immunoprecipitation assays identify IC-binding partners....
Kinesin II is a heterotrimeric plus end–directed microtubule motor responsible for the anterograde movement of organelles in various cell types. Despite substantial literature concerning types that kinesin transports, question how this associates with cargo remains unanswered. To address question, we have used Xenopus laevis melanophores as model system. Through analysis II–mediated melanosome motility, determined dynactin complex, known an anchor cytoplasmic dynein, also links to...
The complete nucleotide sequence of Saccharomyces cerevisiae chromosome VIII reveals that it contains 269 predicted or known genes (300 base pairs larger). Fifty-nine these (22 percent) were previously identified. Of the 210 novel genes, 65 are to encode proteins similar other function. Sixteen appear be relatively recently duplicated. On average, there is one gene approximately every 2 kilobases. Although coding density and composition across not uniform, no regular pattern variation apparent.
A subset of microtubule-associated proteins, including cytoplasmic linker protein (CLIP)-170, dynactin, EB1, adenomatous polyposis coli, dynein, CLASPs, and LIS-1, has been shown recently to target the plus ends microtubules. The mechanisms functions this binding specificity are not understood, although a role in encouraging microtubule elongation proposed. To extend previous work on dynactin organelle transport, we analyzed p150Glued by live-cell imaging. Time-lapse analysis revealed...
ABSTRACT Cytoplasmic dynein is a minus end-directed microtubule motor responsible for centripetal organelle movement and several aspects of chromosome segregation. Our search cytoplasmic dynein-interacting proteins has implicated the dynactin complex as ‘receptor’ on organelles kinetochores. Immunofluorescence microscopy using total six antibodies generated against p150Glued, Arp1 dynamitin subunits revealed novel fraction dynactin-positive structures aligned in linear arrays along distal...
A common feature shared by myosin-binding proteins from a wide variety of species is the presence variable number related internal motifs homologous to either Ig C2 or fibronectin (Fn) type III repeats. Despite interest in potential function these motifs, no group has clearly demonstrated for sequences muscle, intra- extracellularly. We have completed nucleotide sequence fast isoform MyBP-C (C protein) chicken skeletal muscle. The deduced amino acid reveals seven sets and three Fn MyBP-C....
EB1 is a microtubule tip–associated protein that interacts with the APC tumor suppressor and components of dynein/dynactin complex. We have found C-terminal 50 84 amino acids (aa) were sufficient to mediate interactions dynactin, respectively. formed mutually exclusive complexes direct interaction between p150 Glued was identified. EB1-GFP deletion mutants demonstrated role for N-terminus in mediating EB1-microtubule interaction, whereas regions contributed both its tip localization...
A variety of names has been used in the literature for subunits cytoplasmic dynein complexes. Thus, there is a strong need more definitive consensus statement on nomenclature. This especially important mammalian dyneins, many which are encoded by multiple genes. We propose subunit genes and proteins that reflect phylogenetic relationships published studies clarifying functions polypeptides. nomenclature recognizes two distinct complexes flexibility to accommodate discovery new isoforms.
To express the function encoded in its genome, herpes simplex virus 1 capsid-tegument structure released by deenvelopment during entry into cells must be transported retrograde to nuclear pore where viral DNA is nucleus. This path essential case of entering axons dorsal root ganglia. The objective study was identify proteins that may involved transport. We report following findings. (i) neuronal isoform intermediate chain (IC-1a) dynein complex pulled down, from lysates...
Cytoplasmic dynein functions at several sites during mitosis; however, the basis of targeting to each site remains unclear. Tandem mass spectrometry analysis mitotic revealed a phosphorylation in intermediate chains (ICs) that mediates binding kinetochores. IC directs zw10 rather than dynactin, and this interaction is needed for kinetochore localization. Phosphodynein associates with kinetochores from nuclear envelope breakdown metaphase, but bioriented microtubule (MT) attachment chromosome...
Previously, we identified dynactin as a cargo receptor or adaptor for cytoplasmic dynein, mediated by an interaction between the dynein intermediate chain and p150<sup>Glued</sup>. To test phosphorylation potential regulatory mechanism this interaction, analyzed two-dimensional gel analysis detected two variants, one of which was eliminated phosphatase treatment. Overlay assays demonstrated that p150<sup>Glued</sup> bound dephosphorylated but not phosphorylated chains. We then subjected...
Myosin‐binding‐protein C (MyBP‐C) is a myosin‐associated protein of unknown function found in the cross‐bridge‐bearing zone (C region) A bands striated muscle. Using cDNA clone encoding fast‐type isoform chicken MyBP‐C, we screened human fetal muscle library and isolated clones full‐length MyBP‐C. slow‐type were also fully sequenced. Northern‐blot analysis demonstrated skeletal muscle‐specific expression these gene products. human/hamster somatic‐cell hybrids, able to map MyBP‐C chromosome...
Cytoplasmic dynein is one of the major motor proteins involved in intracellular transport. It a protein complex consisting four subunit classes: heavy chains, intermediate chains (ICs), light and chains. In previous study, we had generated new monoclonal antibodies to ICs mapped base motor. Because have been implicated targeting cargo, tested whether these chain could block function cytoplasmic dynein. When extracts Xenopus oocytes were incubated with either (m74-1, m74-2), neither organelle...
The 74-kDa intermediate chains (IC74) of the cytoplasmic dynein complex are believed to be involved in association with membranous organelles. While each molecule is thought have two or three IC74 subunits, at least six different protein isoforms were found from brain. Therefore we investigated relationships brain and their cellular level. We that cultured cortical neurons glia express distinct isoforms. isoform pattern observed was generally similar adult brain, indicating there populations...
The complete nucleotide sequence of a cDNA clone encoding the chicken skeletal muscle myosin-binding protein H (MyBP-H), formerly termed 86-kDa protein, has been established and predicted amino acid compared with other proteins entered into GenBank data base. full-length 2066 base pairs contains single open reading frame 1611 muscle-specific 58,487 Da. molecular weight differs significantly from relative mobility in reducing sodium dodecyl sulfate-polyacrylamide gels (SDS-PAGE). expressed...
Abstract Taxol functions to suppress the dynamic behavior of individual microtubules, and induces multipolar mitotic spindles. However, little is known about mechanisms by which taxol disrupts normal bipolar spindle assembly in vivo. Using live imaging GFP‐α tubulin expressing cells, we examined after treatment. We find that as taxol‐treated cells enter mitosis, there a dramatic re‐distribution microtubule network from centrosomes cell cortex. As they align there, cortical microtubules...
Aurora B (AurB) is a mitotic kinase responsible for multiple aspects of progression, including assembly the outer kinetochore. Cytoplasmic dynein an abundant kinetochore protein whose recruitment to kinetochores requires phosphorylation. To assess whether AurB regulates kinetochores, we inhibited using ZM447439 or kinase-dead construct. Inhibition reduced accumulation at substantially; however, this reflected loss dynein-associated proteins rather than defect in We determined that inhibition...