A5086287356- Cardiomyopathy and Myosin Studies
- Cellular Mechanics and Interactions
- Muscle Physiology and Disorders
- Fish Ecology and Management Studies
- Microtubule and mitosis dynamics
- Genetic diversity and population structure
- Force Microscopy Techniques and Applications
- Fish Biology and Ecology Studies
- Identification and Quantification in Food
- RNA Research and Splicing
- Cellular transport and secretion
- Nuclear Structure and Function
- Developmental Biology and Gene Regulation
- Ion channel regulation and function
- Genetic and Kidney Cyst Diseases
- Genetic and phenotypic traits in livestock
- Trypanosoma species research and implications
- Plant Molecular Biology Research
- Aquatic Invertebrate Ecology and Behavior
- Protist diversity and phylogeny
- Fungal and yeast genetics research
- Water Quality Monitoring Technologies
- Genetics, Aging, and Longevity in Model Organisms
- Water Quality and Pollution Assessment
- Water Quality Monitoring and Analysis
National Heart Lung and Blood Institute
2015-2024
National Institutes of Health
2014-2024
West Virginia University
2023-2024
University of Leeds
2008-2014
National Cancer Institute
2013
Laboratory of Molecular Genetics
2013
Institute of Structural and Molecular Biology
2008-2011
Troy University
2004-2010
Southern Illinois University Carbondale
1992-1998
University of Illinois Urbana-Champaign
1996
Watching proteins' weight Careful measurements of light scattering can provide information on individual macromolecules and complexes. Young et al. used a light-scattering approach for accurate mass determination proteins as small 20 kDa (see the Perspective by Lee Klenerman). Movies protein complex association dissociation were analyzed to extract biophysical parameters from single molecules assemblies without labeling. Using this approach, authors determined in vitro kinetics fibril...
Molecular motors employ chemical energy to generate unidirectional mechanical output against a track while navigating chaotic cellular environment, potential disorder on the track, and Brownian motion. Nevertheless, decades of nanometer-precise optical studies suggest that myosin-5a, one prototypical molecular motors, takes uniform steps spanning 13 subunits (36 nm) along its F-actin track. Here, we use high-resolution interferometric scattering microscopy reveal myosin strides 22 34 actin...
Patterns of mitochondrial DNA (mtDNA) variation have now been examined in more than 40 fish species. This work has established that genomes show constrained size variation, but closely related species often vary size, suggesting counterbalancing deletions and insertions act to stabilize genome size. Gene order composition are also stable, nucleotide sequences rapid divergence. Populations most contain a few common mtDNA haplotypes spectrum their rare mutational derivatives. The extent...
Optical detection of individual proteins requires fluorescent labeling. Cavity and plasmonic methodologies enhance single molecule signatures in the absence any labels but have struggled to demonstrate routine quantitative protein detection. Here, we used interferometric scattering microscopy not only detect also image nanometrically track motion myosin 5a heavy meromyosin molecules without use or nanoscopic amplification. Together with simple experimental arrangement, an intrinsic...
Fluorescent nanodiamonds (FNDs) emit in the near-IR and do not photobleach or photoblink. These properties make FNDs better suited for numerous imaging applications compared with commonly used fluorescence agents such as organic dyes quantum dots. However, form stable suspensions aqueous buffer, are prone to aggregation, difficult functionalize. Here we present a method encapsulating silica using an innovative liposome-based encapsulation process that renders particle surface biocompatible,...
We investigated the role of full-length Drosophila Bicaudal D (BicD) binding partners in dynein-dynactin activation for mRNA transport on microtubules. Full-length BicD robustly activated motility only when both protein Egalitarian (Egl) and K10 cargo were present, electron microscopy showed that Egl needed to disrupt a looped, auto-inhibited conformation. can recruit two dimeric dyneins, resulting faster speeds longer runs than with one dynein. Moving complexes predominantly contained...
Unconventional myosin 15 is a molecular motor expressed in inner ear hair cells that transports protein cargos within developing mechanosensory stereocilia. Mutations of cause profound hearing loss humans and mice; however, the properties this its regulation stereocilia organelle are unknown. To address these questions, we subfragment 1-like (S1) truncation mouse 15, comprising predicted domain plus three light-chain binding sites. Following unsuccessful attempts to express functional 15-S1...
Nonmusclemyosin 2 (NM-2) powers cell motility and tissue morphogenesis by assembling into bipolar filaments that interact with actin. Although the enzymatic properties of purified NM-2 motor fragments have been determined, emergent filament ensembles are unknown. Using single myosin in vitro assays, we report fundamental differences formed different motors. Filaments consisting NM2-B moved processively along actin, while under identical conditions, NM2-A did not. By more closely mimicking...
Local accumulation of oskar (osk) mRNA in the Drosophila oocyte determines posterior pole future embryo. Two major cytoskeletal components, microtubules and actin filaments, together with a microtubule motor, kinesin-1, an myosin-V, are essential for osk localization. In this study, we use Staufen, RNA-binding protein that colocalizes mRNA, as proxy mRNA. We demonstrate localization osk/Staufen is determined by competition between kinesin-1 myosin-V. While removes from cortex along...
Mitochondrial DNA (mtDNA) was isolated from five species of the genus Salvelinus (alpinus, confluentus, fontinalis, malma, and namaycush) closely related taxon, Hucho hucho. Restriction sites recognized by 14 endonucleases were mapped for each percent sequence divergence estimated between pairs. A phylogeny Savelinus based on restriction analysis mtDNA, using as an outgroup, readily resolved three subgenera: Baione, Cristivomer, Salvelinus. Within no differences detected mtDNA brook (S....
Mitochondrial DNA variation was examined in 141 walleye (Stizostedion vitreum) from 10 populations the Great Lakes basin. Twenty-two hexanucleotide sequence recognition endonucleases were used, of which six (Ava I, Bst EII, Cla Dra Nco and Sca I) revealed polymorphisms between fish. Nine mitochondrial clones resolved fell into two major groups that differed their I fragment patterns. The "A" group predominates eastern Lakes, while "B" west. geographical distribution these suggests persisted...
The myosin 2 family of molecular motors includes isoforms regulated in different ways. Vertebrate smooth-muscle is activated by phosphorylation the regulatory light chain, whereas scallop striated adductor-muscle direct calcium binding to its essential chain. paired heads inhibited molecules from myosins have an asymmetric arrangement with motor-motor interactions. It was unknown whether such interactions were a common motif for inactivation used other forms myosin-linked regulation. Using...
The Mus musculus myosin-18A gene is expressed as two alternatively spliced isoforms, α and β, with reported roles in Golgi localization, maintenance of cytoskeleton, receptors for immunological surfactant proteins. Both isoforms feature a myosin motor domain, single predicted IQ motif, long coiled-coil reminiscent myosin-2. myosin-18Aα isoform, additionally, has an N-terminal PDZ domain. Recombinant heavy meromyosin- subfragment-1 (S1)-like constructs both -18β species were purified from the...
The role of nonmuscle myosin 2 (NM2) pulsatile dynamics in generating contractile forces required for developmental morphogenesis has been characterized, but whether these contractions are an intrinsic property all actomyosin networks is not known. Here we used live-cell fluorescence imaging to show that transient, local assembly NM2A “pulses” occurs the cortical cytoskeleton single adherent cells mesenchymal, epithelial, and sarcoma origin, independent signaling cues cell–cell or cell–ECM...
Non-muscle cell contractility is critical for tissues to adopt shape changes. Although, the non-muscle myosin II holoenzyme (myosin) a molecular motor that powers contraction of actin cytoskeleton networks, recent studies have questioned importance activity and tissue Here, combining biochemical analysis enzymatic motile properties purified mutants with in vivo measurements apical constriction same mutants, we show rate scales activity. We so-called phosphomimetic Drosophila regulatory light...
This study has examined the potential use of mitochondrial DNA (mtDNA) variation as a tool for identifying and managing stocks walleye Stizostedion vitreum. Using 10 informative restriction endonucleases, we mtDNA in 847 walleyes from 68 populations across species' range. Thirty-four haplotypes were recognized, 21 which observed only single fish. Eighty-nine percent sampled members one three major haplotypes. These showed distinct geographic distributions: haplotype 1 (12% all fish)...
Twelve populations of walleye (Stizostedion vitreum) from the Great Lakes and three northern Manitoba were screened for allozyme mitochondrial DNA (mtDNA) variation. Nine enzyme loci known to show genetic variation in all fish: five them (Prot-4, Prot-2, Mdh-3, Idh-1, Adh) showed appreciable polymorphism. MtDNA was examined fish using six endonucleases that detected polymorphic sites a further 13 only monomorphic sites. Only one (Prot-4) evidence geographic patterning allele frequencies. By...
Significance Filopodia act as organelles for sensing and exploring the environment, well producing traction forces during cellular locomotion. Myosin-10 is a molecular motor crucial intrafilopodial trafficking filopodia formation. To decipher how myosin-10 generates force movement, we used electron microscopy combination of ensemble biochemical single molecule mechanical techniques to help elucidate its structure mechano-chemical coupling. Our results clarify current controversies about...