'The molecular nature of store-operated Ca(2+)-selective channels has remained an enigma, due largely to the continued inability to convincingly demonstrate Ca(2+)-selective store-operated currents resulting from exogenous expression of known genes. Recent findings have implicated two proteins, Stim1 and Orai1, as having essential roles in store-operated Ca(2+) entry across the plasma ... More
Human stem cell delivery for treatment of large segmental bone defects.
AuthorsDupont KM, Sharma K, Stevens HY, Boerckel JD, García AJ, Guldberg RE,
JournalProc Natl Acad Sci U S A
PubMed ID20133731
'Local or systemic stem cell delivery has the potential to promote repair of a variety of damaged or degenerated tissues. Although various stem cell sources have been investigated for bone repair, few comparative reports exist, and cellular distribution and viability postimplantation remain key issues. In this study, we quantified the ... More
'Pseudogout is an autoinflammatory condition triggered by calcium pyrophosphate dehydrate (CPPD) crystal deposition in the joints. The innate immune system is irritated by and responds to the presence of the crystals with an inflammatory response. The synovial fluid contains activated inflammatory macrophages and neutrophil granulocytes. Several details of crystal-induced macrophage ... More
Androgen induces expression of the multidrug resistance protein gene MRP4 in prostate cancer cells.
AuthorsCai C, Omwancha J, Hsieh CL, Shemshedini L
JournalProstate Cancer Prostatic Dis
PubMed ID17003774
'Multidrug resistance-associated proteins (MRPs) may mediate multidrug resistance in tumor cells. Using a gene array analysis, we have identified MRP4 as an androgen receptor (AR)-regulated gene. Dihydrotestosterone induced MRP4 expression in both androgen-dependent and -independent LNCaP cells, whereas there was little detectable expression in PC-3 or normal prostate epithelial cells. ... More
Estimating relative carbonyl levels in muscle microstructures by fluorescence imaging.
AuthorsFeng J, Navratil M, Thompson LV, Arriaga EA,
JournalAnal Bioanal Chem
PubMed ID18548236
'The increase in the levels of protein carbonyls, biomarkers of oxidative stress, appears to play an important role in aging skeletal muscle. However, the exact distributions of carbonyls among various skeletal muscle microstructures still remain largely unknown, partly owing to the lack of adequate techniques to carry out these measurements. ... More
Three-dimensional resolution doubling in wide-field fluorescence microscopy by structured illumination.
AuthorsGustafsson MG, Shao L, Carlton PM, Wang CJ, Golubovskaya IN, Cande WZ, Agard DA, Sedat JW,
JournalBiophys J
PubMed ID18326650
'Structured illumination microscopy is a method that can increase the spatial resolution of wide-field fluorescence microscopy beyond its classical limit by using spatially structured illumination light. Here we describe how this method can be applied in three dimensions to double the axial as well as the lateral resolution, with true ... More
Integrin alpha5/beta1 expression mediates HER-2 down-regulation in colon cancer cells.
AuthorsKuwada SK, Kuang J, Li X
JournalJ Biol Chem
PubMed ID15757908
'HER-2 is constitutively activated and overexpressed in many cancers, and its inhibition in colon cancer cells diminishes tumorigenicity and induces apoptosis. Little is known about the regulation of HER-2 signaling in colon cancer cells. Integrin alpha5/beta1 expression is frequently lost in colorectal cancer cells compared with normal intestinal epithelium, and ... More
Roles of glutamate receptors and the mammalian target of rapamycin (mTOR) signaling pathway in activity-dependent dendritic protein synthesis in hippocampal neurons.
AuthorsGong R, Park CS, Abbassi NR, Tang SJ
JournalJ Biol Chem
PubMed ID16651266
'Local protein synthesis in neuronal dendrites is critical for synaptic plasticity. However, the signaling cascades that couple synaptic activation to dendritic protein synthesis remain elusive. The purpose of this study is to determine the role of glutamate receptors and the mammalian target of rapamycin (mTOR) signaling in regulating dendritic protein ... More
RANK overexpression in transgenic mice with mouse mammary tumor virus promoter-controlled RANK increases proliferation and impairs alveolar differentiation in the mammary epithelia and disrupts lumen formation in cultured epithelial acini.
AuthorsGonzalez-Suarez E, Branstetter D, Armstrong A, Dinh H, Blumberg H, Dougall WC,
JournalMol Cell Biol
PubMed ID17145767
'RANK and RANKL, the key regulators of osteoclast differentiation and activation, also play an important role in the control of proliferation and differentiation of mammary epithelial cells during pregnancy. Here, we show that RANK protein expression is strictly regulated in a spatial and temporal manner during mammary gland development. RANK ... More
Vascular lipid accumulation, lipoprotein oxidation, and macrophage lipid uptake in hypercholesterolemic zebrafish.
AuthorsStoletov K, Fang L, Choi SH, Hartvigsen K, Hansen LF, Hall C, Pattison J, Juliano J, Miller ER, Almazan F, Crosier P, Witztum JL, Klemke RL, Miller YI,
JournalCirc Res
PubMed ID19265037
'Lipid accumulation in arteries induces vascular inflammation and atherosclerosis, the major cause of heart attack and stroke in humans. Extreme hyperlipidemia induced in mice and rabbits enables modeling many aspects of human atherosclerosis, but microscopic examination of plaques is possible only postmortem. Here we report that feeding adult zebrafish (Danio ... More
TGF-beta1 + EGF-initiated invasive potential in transformed human keratinocytes is coupled to a plasmin/MMP-10/MMP-1-dependent collagen remodeling axis: role for PAI-1.
AuthorsWilkins-Port CE, Ye Q, Mazurkiewicz JE, Higgins PJ,
JournalCancer Res
PubMed ID19383899
'The phenotypic switching called epithelial-to-mesenchymal transition is frequently associated with epithelial tumor cell progression from a comparatively benign to an aggressive, invasive malignancy. Coincident with the emergence of such cellular plasticity is an altered response to transforming growth factor-beta (TGF-beta) as well as epidermal growth factor (EGF) receptor amplification. TGF-beta ... More
Human benign prostatic hyperplasia stromal cells as inducers and targets of chronic immuno-mediated inflammation.
AuthorsPenna G, Fibbi B, Amuchastegui S, Cossetti C, Aquilano F, Laverny G, Gacci M, Crescioli C, Maggi M, Adorini L,
JournalJ Immunol
PubMed ID19299703
'Benign prostatic hyperplasia (BPH), a highly prevalent prostatic condition, could involve an inflammatory component in disease pathogenesis. In this study, we show that human stromal prostate cells obtained from BPH tissue can actively contribute to the inflammatory process by secreting proinflammatory cytokines as well as chemokines able to recruit lymphomonuclear ... More
Spatial distribution and functional significance of activated vinculin in living cells.
AuthorsChen H, Cohen DM, Choudhury DM, Kioka N, Craig SW
JournalJ Cell Biol
PubMed ID15883197
'Conformational change is believed to be important to vinculin''s function at sites of cell adhesion. However, nothing is known about vinculin''s conformation in living cells. Using a Forster resonance energy transfer probe that reports on changes in vinculin''s conformation, we find that vinculin is in the actin-binding conformation in a ... More
Interleukin-15 rescues tolerant CD8+ T cells for use in adoptive immunotherapy of established tumors.
'CD8+ T cells can mediate eradication of established tumors, and strategies to amplify tumor-reactive T-cell numbers by immunization or ex vivo expansion followed by adoptive transfer are currently being explored in individuals with cancer. Generating effective CD8+ T cell-mediated responses to tumors is often impeded by T-cell tolerance to relevant ... More
Regulation of CD4 expression via recycling by HRES-1/RAB4 controls susceptibility to HIV infection.
AuthorsNagy G, Ward J, Mosser DD, Koncz A, Gergely P, Stancato C, Qian Y, Fernandez D, Niland B, Grossman CE, Telarico T, Banki K, Perl A
JournalJ Biol Chem
PubMed ID16935861
'A novel 2986-base transcript encoded by the antisense strand of the HRES-1 human endogenous retrovirus was isolated from peripheral blood lymphocytes. This transcript codes for a 218-amino acid protein, termed HRES-1/Rab4, based on homology to the Rab4 family of small GTPases. Antibody 13407 raised against recombinant HRES-1/Rab4 detected a native ... More
Analysis of ryanodine receptor clusters in rat and human cardiac myocytes.
Single rat ventricular myocytes and human ventricle tissue sections were labeled with antibodies against the ryanodine receptor (RyR) and alpha-actinin to examine the 3D distribution of RyRs with confocal microscopy. Image contrast was maximized by refractive index matching and deconvolution. The RyR label formed discrete puncta representing clusters of RyRs ... More
Visualization and identification of IL-7 producing cells in reporter mice.
AuthorsMazzucchelli RI, Warming S, Lawrence SM, Ishii M, Abshari M, Washington AV, Feigenbaum L, Warner AC, Sims DJ, Li WQ, Hixon JA, Gray DH, Rich BE, Morrow M, Anver MR, Cherry J, Naf D, Sternberg LR, McVicar DW, Farr AG, Germain RN, Rogers K, Jenkins NA, Copeland NG, Durum SK,
JournalPLoS One
PubMed ID19907640
Interleukin-7 (IL-7) is required for lymphocyte development and homeostasis although the actual sites of IL-7 production have never been clearly identified. We produced a bacterial artificial chromosome (BAC) transgenic mouse expressing ECFP in the Il7 locus. The construct lacked a signal peptide and ECFP (enhanced cyan fluorescent protein) accumulated inside ... More
Cloning and pharmacological characterization of the guinea pig P2X7 receptor orthologue.
AuthorsFonfria E, Clay WC, Levy DS, Goodwin JA, Roman S, Smith GD, Condreay JP, Michel AD,
JournalBr J Pharmacol
PubMed ID18037910
BACKGROUND AND PURPOSE: The human, rat, and mouse P2X(7) receptors have been previously characterized, and in this study we report the cloning and pharmacological properties of the guinea pig orthologue. EXPERIMENTAL APPROACH: A cDNA encoding for the guinea pig P2X(7) receptor was isolated from a guinea pig brain library. The ... More
Transcriptional repression mediated by repositioning of genes to the nuclear lamina.
AuthorsReddy KL, Zullo JM, Bertolino E, Singh H,
JournalNature
PubMed ID18272965
Nuclear compartmentalization seems to have an important role in regulating metazoan genes. Although studies on immunoglobulin and other loci have shown a correlation between positioning at the nuclear lamina and gene repression, the functional consequences of this compartmentalization remain untested. We devised an approach for inducible tethering of genes to ... More
Identification of a small GTPase inhibitor using a high-throughput flow cytometry bead-based multiplex assay.
AuthorsSurviladze Z, Waller A, Wu Y, Romero E, Edwards BS, Wandinger-Ness A, Sklar LA,
JournalJ Biomol Screen
PubMed ID20008126
Small GTPases are key regulators of cellular activity and represent novel targets for the treatment of human diseases using small-molecule inhibitors. The authors describe a multiplex, flow cytometry bead-based assay for the identification and characterization of inhibitors or activators of small GTPases. Six different glutathione-S-transferase (GST)-tagged small GTPases were bound ... More
DAG lipase activity is necessary for TRP channel regulation in Drosophila photoreceptors.
AuthorsLeung HT, Tseng-Crank J, Kim E, Mahapatra C, Shino S, Zhou Y, An L, Doerge RW, Pak WL,
JournalNeuron
PubMed ID18579079
In Drosophila, a phospholipase C-mediated signaling cascade links photoexcitation of rhodopsin to the opening of the TRP/TRPL channels. A lipid product of the cascade, diacylglycerol (DAG) and its metabolite(s), polyunsaturated fatty acids (PUFAs), have both been proposed as potential excitatory messengers. A crucial enzyme in the understanding of this process ... More
LytA, major autolysin of Streptococcus pneumoniae, requires access to nascent peptidoglycan.
The pneumococcal autolysin LytA is a virulence factor involved in autolysis as well as in fratricidal- and penicillin-induced lysis. In this study, we used biochemical and molecular biological approaches to elucidate which factors control the cytoplasmic translocation and lytic activation of LytA. We show that LytA is mainly localized intracellularly, ... More
NS21: Re-defined and modified supplement B27 for neuronal cultures.
In vitro culturing of primary neurons is a mainstay of neurobiological research. Many of these culture paradigms have taken advantage of defined culture media rather than serum additives that contain undefined survival factors to facilitate experimental manipulations and interpretation of the results. To culture neurons in the absence of serum, ... More
Novel pentablock copolymers for selective gene delivery to cancer cells.
AuthorsZhang B, Kanapathipillai M, Bisso P, Mallapragada S,
JournalPharm Res
PubMed ID19142716
PURPOSE: In this study, the novel poly(diethylaminoethylmethacrylate) (PDEAEM)/Pluronic F127 pentablock copolymers were found to be able to mediate high-efficiency transfection of human epithelial ovarian carcinoma (SKOV3) cell line while showing significantly lower efficacy in human epithelial retinal (ARPE-19) cell line and Swiss Mouse Fibroblast (3T3) cell line. METHODS: The intracellular ... More
Heat sensitization in skin and muscle nociceptors expressing distinct combinations of TRPV1 and TRPV2 protein.
AuthorsRau KK, Jiang N, Johnson RD, Cooper BY,
JournalJ Neurophysiol
PubMed ID17287441
Recordings were made from small and medium diameter dorsal root ganglia (DRG) neurons that expressed transient receptor potential (TRP) proteins. Physiologically characterized skin nociceptors expressed either TRPV1 (type 2) or TRPV2 (type 4) in isolation. Other nociceptors co-expressed both TRP proteins and innervated deep tissue sites (gastrocnemius muscle, distal colon; ... More
In situ visualization and dynamics of newly synthesized proteins in rat hippocampal neurons.
AuthorsDieterich DC, Hodas JJ, Gouzer G, Shadrin IY, Ngo JT, Triller A, Tirrell DA, Schuman EM,
JournalNat Neurosci
PubMed ID20543841
Protein translation has been implicated in different forms of synaptic plasticity, but direct in situ visualization of new proteins is limited to one or two proteins at a time. Here we describe a metabolic labeling approach based on incorporation of noncanonical amino acids into proteins followed by chemoselective fluorescence tagging ... More
Quantitative fluorescence imaging analysis for cancer biomarker discovery: application to beta-catenin in archived prostate specimens.
AuthorsHuang D, Casale GP, Tian J, Wehbi NK, Abrahams NA, Kaleem Z, Smith LM, Johansson SL, Elkahwaji JE, Hemstreet GP,
JournalCancer Epidemiol Biomarkers Prev
PubMed ID17623804
The surprising disparity between the number of protein-encoding genes ( approximately 30,000) in the human genome and the number of proteins ( approximately 300,000) in the human proteome has inspired the development of translational proteomics aimed at protein expression profiling of disease states. Translational proteomics, which offers the promise of ... More
Dendritic cell-nerve clusters are sites of T cell proliferation in allergic airway inflammation.
AuthorsVeres TZ, Shevchenko M, Krasteva G, Spies E, Prenzler F, Rochlitzer S, Tschernig T, Krug N, Kummer W, Braun A,
JournalAm J Pathol
PubMed ID19179611
Interactions between T cells and dendritic cells in the airway mucosa precede secondary immune responses to inhaled antigen. The purpose of this study was to identify the anatomical locations where dendritic cell-T cell interactions occur, resulting in T cells activation by dendritic cells. In a mouse model of allergic airway ... More
A stochastic mechanism for biofilm formation by Mycoplasma pulmonis.
Bacterial biofilms are communities of bacteria that are enclosed in an extracellular matrix. Within a biofilm the bacteria are protected from antimicrobials, environmental stresses, and immune responses from the host. Biofilms are often believed to have a highly developed organization that is derived from differential regulation of the genes that ... More
Live-cell superresolution imaging by pulsed STED two-photon excitation microscopy.
AuthorsTakasaki KT, Ding JB, Sabatini BL,
JournalBiophys J
PubMed ID23442955
Two-photon laser scanning microscopy (2PLSM) allows fluorescence imaging in thick biological samples where absorption and scattering typically degrade resolution and signal collection of one-photon imaging approaches. The spatial resolution of conventional 2PLSM is limited by diffraction, and the near-infrared wavelengths used for excitation in 2PLSM preclude the accurate imaging of ... More
The transmembrane domain of the severe acute respiratory syndrome coronavirus ORF7b protein is necessary and sufficient for its retention in the Golgi complex.
AuthorsSchaecher SR, Diamond MS, Pekosz A,
JournalJ Virol
PubMed ID18632859
The severe acute respiratory syndrome coronavirus (SARS-CoV) ORF7b (also called 7b) protein is an integral membrane protein that is translated from a bicistronic open reading frame encoded within subgenomic RNA 7. When expressed independently or during virus infection, ORF7b accumulates in the Golgi compartment, colocalizing with both cis- and trans-Golgi ... More
Robust fluorescent detection of protein fatty-acylation with chemical reporters.
AuthorsCharron G, Zhang MM, Yount JS, Wilson J, Raghavan AS, Shamir E, Hang HC,
JournalJ Am Chem Soc
PubMed ID19281244
Fatty-acylation of proteins in eukaryotes is associated with many fundamental cellular processes but has been challenging to study due to limited tools for rapid and robust detection of protein fatty-acylation in cells. The development of azido-fatty acids enabled the nonradioactive detection of fatty-acylated proteins in mammalian cells using the Staudinger ... More
Microfluidic-based cell sorting of Francisella tularensis infected macrophages using optical forces.
AuthorsPerroud TD, Kaiser JN, Sy JC, Lane TW, Branda CS, Singh AK, Patel KD,
JournalAnal Chem
PubMed ID18510341
We have extended the principle of optical tweezers as a noninvasive technique to actively sort hydrodynamically focused cells based on their fluorescence signal in a microfluidic device. This micro fluorescence-activated cell sorter (microFACS) uses an infrared laser to laterally deflect cells into a collection channel. Green-labeled macrophages were sorted from ... More
A contiguous compartment functions as endoplasmic reticulum and endosome/lysosome in Giardia lamblia.
AuthorsAbodeely M, DuBois KN, Hehl A, Stefanic S, Sajid M, DeSouza W, Attias M, Engel JC, Hsieh I, Fetter RD, McKerrow JH,
JournalEukaryot Cell
PubMed ID19749174
The dynamic evolution of organelle compartmentalization in eukaryotes and how strictly compartmentalization is maintained are matters of ongoing debate. While the endoplasmic reticulum (ER) is classically envisioned as the site of protein cotranslational translocation, it has recently been proposed to have pluripotent functions. Using transfected reporter constructs, organelle-specific markers, and ... More
Homologous recombinational repair factors are recruited and loaded onto the viral DNA genome in Epstein-Barr virus replication compartments.
AuthorsKudoh A, Iwahori S, Sato Y, Nakayama S, Isomura H, Murata T, Tsurumi T,
JournalJ Virol
PubMed ID19386720
Homologous recombination is an important biological process that facilitates genome rearrangement and repair of DNA double-strand breaks (DSBs). The induction of Epstein-Barr virus (EBV) lytic replication induces ataxia telangiectasia-mutated (ATM)-dependent DNA damage checkpoint signaling, leading to the clustering of phosphorylated ATM and Mre11/Rad50/Nbs1 (MRN) complexes to sites of viral genome ... More
Metabolic installation of thiols into sialic acid modulates adhesion and stem cell biology.
AuthorsSampathkumar SG, Li AV, Jones MB, Sun Z, Yarema KJ
JournalNat Chem Biol
PubMed ID16474386
Metabolic 'oligosaccharide engineering' methods based on N-acetyl-D-mannosamine (ManNAc) analogs allow the glycocalyx of living cells to be remodeled. Herein we report the analog Ac(5)ManNTGc (1) that enables thiols to be expressed in surface sialic acids. By locating this versatile functional group on the outer periphery of normally nonadhesive human Jurkat ... More
gp130 signaling in proopiomelanocortin neurons mediates the acute anorectic response to centrally applied ciliary neurotrophic factor.
Ciliary neurotrophic factor (CNTF) exerts anorectic effects by overcoming leptin resistance via activation of hypothalamic neurons. However, the exact site of CNTF action in the hypothalamus has not yet been identified. Using Cre-loxP-mediated recombination in vivo, we have selectively ablated the common cytokine signaling chain gp130, which is required for ... More
A mutant alphaII-spectrin designed to resist calpain and caspase cleavage questions the functional importance of this process in vivo.
AuthorsMeary F, Metral S, Ferreira C, Eladari D, Colin Y, Lecomte MC, Nicolas G
JournalJ Biol Chem
PubMed ID17374614
alpha- and beta-spectrins are components of molecular scaffolds located under the lipid bilayer and named membrane skeletons. Disruption of these scaffolds through mutations in spectrins demonstrated that they are involved in the membrane localization or the maintenance of proteins associated with them. The ubiquitous alphaII-spectrin chain bears in its central ... More
Phospholipid biosynthesis program underlying membrane expansion during B-lymphocyte differentiation.
AuthorsFagone P, Sriburi R, Ward-Chapman C, Frank M, Wang J, Gunter C, Brewer JW, Jackowski S
JournalJ Biol Chem
PubMed ID17213195
Stimulated B-lymphocytes differentiate into plasma cells committed to antibody production. Expansion of the endoplasmic reticulum and Golgi compartments is a prerequisite for high rate synthesis, assembly, and secretion of immunoglobulins. The bacterial cell wall component lipopolysaccharide (LPS) stimulates murine B-cells to proliferate and differentiate into antibody-secreting cells that morphologically resemble ... More
DNA-binding and transactivation activities are essential for TAp63 protein degradation.
The p53-related p63 gene encodes six isoforms with differing N and C termini. TAp63 isoforms possess a transactivation domain at the N terminus and are able to transactivate a set of genes, including some targets downstream of p53. Accumulating evidence indicates that TAp63 plays an important role in regulation of ... More
Internalization of novel non-viral vector TAT-streptavidin into human cells.
AuthorsRinne J, Albarran B, Jylhävä J, Ihalainen TO, Kankaanpää P, Hytönen VP, Stayton PS, Kulomaa MS, Vihinen-Ranta M
JournalBMC Biotechnol
PubMed ID17199888
BACKGROUND: The cell-penetrating peptide derived from the Human immunodeficiency virus-1 transactivator protein Tat possesses the capacity to promote the effective uptake of various cargo molecules across the plasma membrane in vitro and in vivo. The objective of this study was to characterize the uptake and delivery mechanisms of a novel ... More
Fast noninvasive activation and inhibition of neural and network activity by vertebrate rhodopsin and green algae channelrhodopsin.
AuthorsLi X, Gutierrez DV, Hanson MG, Han J, Mark MD, Chiel H, Hegemann P, Landmesser LT, Herlitze S
JournalProc Natl Acad Sci U S A
PubMed ID16306259
Techniques for fast noninvasive control of neuronal excitability will be of major importance for analyzing and understanding neuronal networks and animal behavior. To develop these tools we demonstrated that two light-activated signaling proteins, vertebrate rat rhodopsin 4 (RO4) and the green algae channelrhodospin 2 (ChR2), could be used to control ... More
Excess Translation of Epigenetic Regulators Contributes to Fragile X Syndrome and Is Alleviated by Brd4 Inhibition.
Authors
JournalCell
PubMed ID28823556
The Golgi Outpost Protein TPPP Nucleates Microtubules and Is Critical for Myelination.
Authors
JournalCell
PubMed ID31522887
Senescence-Induced Vascular Remodeling Creates Therapeutic Vulnerabilities in Pancreas Cancer.
Authors
JournalCell
PubMed ID32234521
Myc Cooperates with Ras by Programming Inflammation and Immune Suppression.
Authors
JournalCell
PubMed ID29195074
Mutant KRAS Enhances Tumor Cell Fitness by Upregulating Stress Granules.
Authors
JournalCell
PubMed ID27984728
Cell Types of the Human Retina and Its Organoids at Single-Cell Resolution.
Authors
JournalCell
PubMed ID32946783
A Membraneless Organelle Associated with the Endoplasmic Reticulum Enables 3'UTR-Mediated Protein-Protein Interactions.
Authors
JournalCell
PubMed ID30449617
Nuclear-Import Receptors Reverse Aberrant Phase Transitions of RNA-Binding Proteins with Prion-like Domains.
Authors
JournalCell
PubMed ID29677512
Mitochondrial Sirtuin Network Reveals Dynamic SIRT3-Dependent Deacetylation in Response to Membrane Depolarization.
Authors
JournalCell
PubMed ID27881304
Circulating Tumor Cell Clustering Shapes DNA Methylation to Enable Metastasis Seeding.