BACKGROUND: Immune checkpoint blockade (ICB) is rapidly becoming a standard of care in the treatment of many cancer types. However, the subset of patients who respond to this type of therapy is limited. Another way to promote antitumoral immunity is the use of immunostimulatory molecules, such as cytokines or T ... More
Mammalian cell penetration, siRNA transfection, and DNA transfection by supercharged proteins.
AuthorsMcNaughton BR, Cronican JJ, Thompson DB, Liu DR,
JournalProc Natl Acad Sci U S A
PubMed ID19307578
'Nucleic acid reagents, including small interfering RNA (siRNA) and plasmid DNA, are important tools for the study of mammalian cells and are promising starting points for the development of new therapeutic agents. Realizing their full potential, however, requires nucleic acid delivery reagents that are simple to prepare, effective across many ... More
Amino acid residues critical for endoplasmic reticulum export and trafficking of platelet-activating factor receptor.
AuthorsHirota N, Yasuda D, Hashidate T, Yamamoto T, Yamaguchi S, Nagamune T, Nagase T, Shimizu T, Nakamura M,
JournalJ Biol Chem
PubMed ID20007715
'Several residues are conserved in the transmembrane domains (TMs) of G-protein coupled receptors. Here we demonstrate that a conserved proline, Pro(247), in TM6 of platelet-activating factor receptor (PAFR) is required for endoplasmic reticulum (ER) export and trafficking after agonist-induced internalization. Alanine-substituted mutants of the conserved residues of PAFRs, including P247A, ... More
Caveolin regulates endocytosis of the muscle repair protein, dysferlin.
AuthorsHernández-Deviez DJ, Howes MT, Laval SH, Bushby K, Hancock JF, Parton RG,
JournalJ Biol Chem
PubMed ID18096699
'Dysferlin and Caveolin-3 are plasma membrane proteins associated with muscular dystrophy. Patients with mutations in the CAV3 gene show dysferlin mislocalization in muscle cells. By utilizing caveolin-null cells, expression of caveolin mutants, and different mutants of dysferlin, we have dissected the site of action of caveolin with respect to dysferlin ... More
Functional analysis of FSP27 protein regions for lipid droplet localization, caspase-dependent apoptosis, and dimerization with CIDEA.
AuthorsLiu K, Zhou S, Kim JY, Tillison K, Majors D, Rearick D, Lee JH, Fernandez-Boyanapalli RF, Barricklow K, Houston MS, Smas CM,
JournalAm J Physiol Endocrinol Metab
PubMed ID19843876
'The adipocyte-specific protein FSP27, also known as CIDEC, is one of three cell death-inducing DFF45-like effector (CIDE) proteins. The first known function for CIDEs was promotion of apoptosis upon ectopic expression in mammalian cells. Recent studies in endogenous settings demonstrated key roles for CIDEs in energy metabolism. FSP27 is a ... More
Loss of PINK1 function promotes mitophagy through effects on oxidative stress and mitochondrial fission.
AuthorsDagda RK, Cherra SJ, Kulich SM, Tandon A, Park D, Chu CT,
JournalJ Biol Chem
PubMed ID19279012
'Mitochondrial dysregulation is strongly implicated in Parkinson disease. Mutations in PTEN-induced kinase 1 (PINK1) are associated with familial parkinsonism and neuropsychiatric disorders. Although overexpressed PINK1 is neuroprotective, less is known about neuronal responses to loss of PINK1 function. We found that stable knockdown of PINK1 induced mitochondrial fragmentation and autophagy ... More
Real-time visualization of HIV-1 GAG trafficking in infected macrophages.
AuthorsGousset K, Ablan SD, Coren LV, Ono A, Soheilian F, Nagashima K, Ott DE, Freed EO,
JournalPLoS Pathog
PubMed ID18369466
'HIV-1 particle production is driven by the Gag precursor protein Pr55(Gag). Despite significant progress in defining both the viral and cellular determinants of HIV-1 assembly and release, the trafficking pathway used by Gag to reach its site of assembly in the infected cell remains to be elucidated. The Gag trafficking ... More
Thiazolides, a new class of anti-influenza molecules targeting viral hemagglutinin at the post-translational level.
AuthorsRossignol JF, La Frazia S, Chiappa L, Ciucci A, Santoro MG,
JournalJ Biol Chem
PubMed ID19638339
'The emergence of highly contagious influenza A virus strains, such as the new H1N1 swine influenza, represents a serious threat to global human health. Efforts to control emerging influenza strains focus on surveillance and early diagnosis, as well as development of effective vaccines and novel antiviral drugs. Herein we document ... More
Random walk of processive, quantum dot-labeled myosin Va molecules within the actin cortex of COS-7 cells.
AuthorsNelson SR, Ali MY, Trybus KM, Warshaw DM,
JournalBiophys J
PubMed ID19619465
'Myosin Va (myoVa) is an actin-based intracellular cargo transporter. In vitro experiments have established that a single myoVa moves processively along actin tracks, but less is known about how this motor operates within cells. Here we track the movement of a quantum dot (Qdot)-labeled myoVa HMM in COS-7 cells using ... More
Inhibiting colorectal carcinoma growth and metastasis by blocking the expression of VEGF using RNA interference.
AuthorsWang S, Liu H, Ren L, Pan Y, Zhang Y,
JournalNeoplasia
PubMed ID18392139
'Angiogenesis plays an essential role in tumor growth and metastasis and is a promising target for cancer therapy. Vascular endothelial growth factor (VEGF) is a key regulator of angiogenesis. The present study was designed to determine the role of VEGF in tumor growth and metastasis using RNA interference (RNAi) technology. ... More
Dual fluorescent labeling method to visualize plasmid DNA degradation.
'The efficiency of nonviral vectors for gene delivery may be enhanced by understanding the key barriers that limit the translocation of the therapeutic DNA into the nucleus. One such barrier is the instability of DNA in the cytoplasm. In this work, we have developed a method to dual-label plasmid DNA ... More
Intensity calibration and automated cell cycle gating for high-throughput image-based siRNA screens of mammalian cells.
AuthorsPoon SS, Wong JT, Saunders DN, Ma QC, McKinney S, Fee J, Aparicio SA,
JournalCytometry A
PubMed ID18698634
'High-content microscopic screening systems are powerful tools for extracting quantitative multiparameter measures from large number of cells under numerous conditions. These systems perform well in applications that monitor the presence of objects, but lack in their ability to accurately estimate object intensities and summarize these findings due to variations in ... More
The cytoplasmic tail dileucine motif LL572 determines the glycosylation pattern of membrane-type 1 matrix metalloproteinase.
'Membrane-type 1 matrix metalloproteinase (MT1-MMP; MMP-14) drives fundamental physiological and pathological processes, due to its ability to process a broad spectrum of substrates. Because subtle changes in its activity can produce profound physiological effects, MT1-MMP is tightly regulated. Currently, many aspects of this regulation remain to be elucidated. It has ... More
The human GINS complex associates with Cdc45 and MCM and is essential for DNA replication.
AuthorsAparicio T, Guillou E, Coloma J, Montoya G, Méndez J,
JournalNucleic Acids Res
PubMed ID19223333
'The GINS complex, originally discovered in Saccharomyces cerevisiae and Xenopus laevis, binds to DNA replication origins shortly before the onset of S phase and travels with the replication forks after initiation. In this study we present a detailed characterization of the human GINS (hGINS) homolog. Using new antibodies that allow ... More
Coculture methodologies for the study of Wnt signals.
AuthorsPlanutis K, Planutiene M, Holcombe RF,
JournalMethods Mol Biol
PubMed ID19099261
In vivo, responses to extracellular Wnt ligands are context dependent; the temporal characteristics and intensity of the signal are critical in determining the target cell response. In general, Wnt ligand-induced differentiation in mammalian cells requires several days of exposure. In order to better characterize Wnt-induced signaling in vitro, side-by-side and ... More
alpha-Actinin interacts with rapsyn in agrin-stimulated AChR clustering.
AuthorsDobbins GC, Luo S, Yang Z, Xiong WC, Mei L,
JournalMol Brain
PubMed ID19055765
: AChR is concentrated at the postjunctional membrane at the neuromuscular junction. However, the underlying mechanism is unclear. We show that alpha-actinin, a protein known to cross-link F-actin, interacts with rapsyn, a scaffold protein essential for neuromuscular junction formation. alpha-Actinin, rapsyn, and surface AChR form a ternary complex. Moreover, the ... More
Esterases in the house fly. Polymorphisms and inheritance patterns.
AuthorsNarang S, Terranova AC, McDonald IC, Leopold RA,
JournalJ Hered
PubMed ID1262698
Polyacrylamide gel electrophoresis was used to examine the variability and inheritance of esterases in five strains of the house fly, Musca domestica L. Individual zymograms exhibited 8 to 15 bands that could be assigned to one of five zones designated as A through E from anode to cathode. Correlations of ... More
Gene transfer of somatostatin receptor type 2 by intratumoral injection inhibits established pancreatic carcinoma xenografts.
AuthorsDu ZY, Qin RY, Xia W, Tian R, Kumar M,
JournalWorld J Gastroenterol
PubMed ID15641137
AIM: To investigate the therapeutic effect of somatostatin receptor type 2 (SSTR2) gene transfection on pancreatic carcinoma xenografts in vivo in experimental cancers. METHODS: Human pancreatic cancer cell line Panc-1 was inoculated subcutaneously into the back of nude mice. When tumor nodules were grown as large as about 5 mmX5 ... More
Regulation of interleukin-8 via an airway epithelial signaling cascade.
Airways function as an innate immune organ against airborne bacteria that are inhaled and deposited in airways. One of the mechanisms of host defense is to recruit neutrophils into airways to clear the invaders. Airway epithelial cells produce neutrophil chemoattractant interleukin (IL)-8 in response to invading bacteria. In this study ... More
Human cytomegalovirus plasmid-based amplicon vector system for gene therapy.
AuthorsMahmood K, Prichard MN, Duke GM, Kemble GW, Spaete RR,
JournalGenet Vaccines Ther
PubMed ID15673469
We have constructed and evaluated the utility of a helper-dependent virus vector system that is derived from Human Cytomegalovirus (HCMV). This vector is based on the herpes simplex virus (HSV) amplicon system and contains the HCMV orthologs of the two cis-acting functions required for replication and packaging of HSV genomes, ... More
Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA.
BACKGROUND: The ability to manipulate the genetic networks underlying the physiological and behavioural repertoires of the adult honeybee worker (Apis mellifera) is likely to deepen our understanding of issues such as learning and memory generation, ageing, and the regulatory anatomy of social systems in proximate as well as evolutionary terms. ... More
An eight residue fragment of an acyl carrier protein suffices for post-translational introduction of fluorescent pantetheinyl arms in protein modification in vitro and in vivo.
AuthorsZhou Z, Koglin A, Wang Y, McMahon AP, Walsh CT,
JournalJ Am Chem Soc
PubMed ID18593165
Genetically encoded tags for tracking a given protein continue to be of great interest in a multitude of in vitro and in vivo contexts. Acyl carrier proteins, both free-standing and as embedded 80-100 residue domains, contain a specific serine side chain that undergoes post-translational pantetheinylation from CoASH as donor substrate. ... More
beta-Galactosidase activity assay using far-red-shifted fluorescent substrate DDAOG.
AuthorsGong H, Zhang B, Little G, Kovar J, Chen H, Xie W, Schutz-Geschwender A, Olive DM,
JournalAnal Biochem
PubMed ID19103143
beta-Galactosidase (beta-gal) is commonly used as a reporter gene in biological research, and a wide variety of substrates have been developed to assay its activity. One substrate, 9H-(1,3-dichloro-9,9-dimethylacridin-2-one-7-yl) beta-d-galactopyranoside (DDAOG), can be cleaved by beta-gal to produce 7-hydroxy-9H(I,3-dichloro-9,9-dimethylacridin-2-one) (DDAO). On excitation, DDAO generates a far-red-shifted fluorescent signal. Using this substrate, ... More
Ankyrin-G is a molecular partner of E-cadherin in epithelial cells and early embryos.
AuthorsKizhatil K, Davis JQ, Davis L, Hoffman J, Hogan BL, Bennett V,
JournalJ Biol Chem
PubMed ID17620337
E-cadherin is a ubiquitous component of lateral membranes in epithelial tissues and is required to form the first lateral membrane domains in development. Here, we identify ankyrin-G as a molecular partner of E-cadherin and demonstrate that ankyrin-G and beta-2-spectrin are required for accumulation of E-cadherin at the lateral membrane in ... More
Amplification of CD95 activation by caspase 8-induced endosomal acidification in rat hepatocytes.
AuthorsReinehr R, Sommerfeld A, Keitel V, Grether-Beck S, Haüssinger D,
JournalJ Biol Chem
PubMed ID18045865
Although in rat hepatocytes CD95 is predominantly located inside the cell with almost undetectable immunostaining at the plasma membrane, the addition of CD95-ligand (CD95L) induces hepatocyte apoptosis, which is preceded by a targeting and activation of intracellularly localized CD95 to the plasma membrane including formation of the death-inducing signaling complex. ... More
Defining mechanisms of actin polymerization and depolymerization during dendritic spine morphogenesis.
Dendritic spines are small protrusions along dendrites where the postsynaptic components of most excitatory synapses reside in the mature brain. Morphological changes in these actin-rich structures are associated with learning and memory formation. Despite the pivotal role of the actin cytoskeleton in spine morphogenesis, little is known about the mechanisms ... More
Automated organelle-based colocalization in whole-cell imaging.
AuthorsWoodcroft BJ, Hammond L, Stow JL, Hamilton NA,
JournalCytometry A
PubMed ID19746416
The use of fluorescence microscopy to investigate protein colocalization is an invaluable tool for understanding subcellular structures and their associated proteins. However, current techniques are largely limited to two-dimensional (2D) imaging and often require manual segmentation. Here, we present OBCOL, a methodology to automatically segment and quantify protein colocalization not ... More
Knockdown of mu-calpain in Fanconi anemia, FA-A, cells by siRNA restores alphaII spectrin levels and corrects chromosomal instability and defective DNA interstrand cross-link repair.
AuthorsZhang P, Sridharan D, Lambert MW,
JournalBiochemistry
PubMed ID20518497
We have previously shown that there is a deficiency in the structural protein, nonerythroid alpha spectrin (alphaIISp), in cells from patients with Fanconi anemia (FA). These studies indicate that this deficiency is due to the reduced stability of alphaIISp and correlates with a decreased level of repair of DNA interstrand ... More
Endogenous 24(S),25-epoxycholesterol fine-tunes acute control of cellular cholesterol homeostasis.
AuthorsWong J, Quinn CM, Gelissen IC, Brown AJ,
JournalJ Biol Chem
PubMed ID17981807
Certain oxysterols, when added to cultured cells, are potent regulators of cholesterol homeostasis, decreasing cholesterol synthesis and uptake and increasing cholesterol efflux. However, very little is known about whether or not endogenous oxysterol(s) plays a significant role in cholesterol homeostasis. 24(S),25-Epoxycholesterol (24,25EC) is unique among oxysterols in that it is ... More
Resveratrol protects mitochondria against oxidative stress through AMP-activated protein kinase-mediated glycogen synthase kinase-3beta inhibition downstream of poly(ADP-ribose)polymerase-LKB1 pathway.
AuthorsShin SM, Cho IJ, Kim SG,
JournalMol Pharmacol
PubMed ID19620254
Arachidonic acid (AA, a proinflammatory fatty acid) in combination with iron promotes excess reactive oxygen species (ROS) production and exerts a deleterious effect on mitochondria. We have shown previously that activation of AMP-activated protein kinase (AMPK) protects hepatocytes from AA + iron-induced apoptosis. Resveratrol, a polyphenol in grapes, has beneficial ... More
Stroma-dependent apoptosis in clonal hematopoietic precursors correlates with expression of PYCARD.
The role of the marrow microenvironment in the pathophysiology of myelodysplastic syndromes (MDSs) remains controversial. Using stromal/hematopoietic cell cocultures, we investigated the effects of stroma-derived signals on apoptosis sensitivity in hematopoietic precursors. The leukemia-derived cell line KG1a is resistant to proapoptotic ligands. However, when cocultured with the human stromal cell ... More
Autophagy in neurite injury and neurodegeneration: in vitro and in vivo models.
Recent advances indicate that maintaining a balanced level of autophagy is critically important for neuronal health and function. Pathologic dysregulation of macroautophagy has been implicated in synaptic dysfunction, cellular stress, and neuronal cell death. Autophagosomes and autolysosomes are induced in acute and chronic neurological disorders including stroke, brain trauma, neurotoxin ... More
S-glutathionylation impairs signal transducer and activator of transcription 3 activation and signaling.
AuthorsXie Y, Kole S, Precht P, Pazin MJ, Bernier M,
JournalEndocrinology
PubMed ID18988672
S-glutathionylation is a physiological, reversible protein modification of cysteine residues with glutathione in response to mild oxidative stress. Because the key cell growth regulator signal transducer and activator of transcription (STAT) 3 is particularly susceptible to redox regulation, we hypothesized that oxidative modification of cysteine residues of STAT3 by S-glutathionylation ... More
Enterocyte TLR4 mediates phagocytosis and translocation of bacteria across the intestinal barrier.
AuthorsNeal MD, Leaphart C, Levy R, Prince J, Billiar TR, Watkins S, Li J, Cetin S, Ford H, Schreiber A, Hackam DJ,
JournalJ Immunol
PubMed ID16493066
Translocation of bacteria across the intestinal barrier is important in the pathogenesis of systemic sepsis, although the mechanisms by which bacterial translocation occurs remain largely unknown. We hypothesized that bacterial translocation across the intact barrier occurs after internalization of the bacteria by enterocytes in a process resembling phagocytosis and that ... More
Mitochondrially localized ERK2 regulates mitophagy and autophagic cell stress: implications for Parkinson's disease.
AuthorsDagda RK, Zhu J, Kulich SM, Chu CT,
JournalAutophagy
PubMed ID18594198
Degenerating neurons of Parkinson's disease (PD) patient brains exhibit granules of phosphorylated extracellular signal-regulated protein kinase 1/2 (ERK1/2) that localize to autophagocytosed mitochondria. Here we show that 6-hydroxydopamine (6-OHDA) elicits activity-related localization of ERK1/2 in mitochondria of SH-SY5Y cells, and these events coincide with induction of autophagy and precede mitochondrial ... More
Hereditary parkinsonism with dementia is caused by mutations in ATP13A2, encoding a lysosomal type 5 P-type ATPase.
AuthorsRamirez A, Heimbach A, Gründemann J, Stiller B, Hampshire D, Cid LP, Goebel I, Mubaidin AF, Wriekat AL, Roeper J, Al-Din A, Hillmer AM, Karsak M, Liss B, Woods CG, Behrens MI, Kubisch C,
JournalNat Genet
PubMed ID16964263
Neurodegenerative disorders such as Parkinson and Alzheimer disease cause motor and cognitive dysfunction and belong to a heterogeneous group of common and disabling disorders. Although the complex molecular pathophysiology of neurodegeneration is largely unknown, major advances have been achieved by elucidating the genetic defects underlying mendelian forms of these diseases. ... More
Functional properties of alpha7 nicotinic acetylcholine receptors co-expressed with RIC-3 in a stable recombinant CHO-K1 cell line.
Heterologous functional expression of alpha7 nicotinic acetylcholine receptors (nAChRs) is difficult to achieve in mammalian cell lines, and the reasons have been associated with a lack of expression of the putative chaperone factor RIC-3. Here, we describe the generation and functional and pharmacological characterization of a Chinese hamster ovary (CHO)-K1 ... More
Dkk-1 inhibits intestinal epithelial cell migration by attenuating directional polarization of leading edge cells.
AuthorsKoch S, Capaldo CT, Samarin S, Nava P, Neumaier I, Skerra A, Sacks DB, Parkos CA, Nusrat A,
JournalMol Biol Cell
PubMed ID19776352
Wnt signaling pathways regulate proliferation, motility, and survival in a variety of human cell types. Dickkopf-1 (Dkk-1) is a secreted Wnt antagonist that has been proposed to regulate tissue homeostasis in the intestine. In this report, we show that Dkk-1 is secreted by intestinal epithelial cells after wounding and that ... More
Differential regulation of macropinocytosis by Abi1/Hssh3bp1 isoforms.
BACKGROUND: Macropinocytosis, which is a constitutive cellular process of fluid and macromolecule uptake, is regulated by actin cytoskeleton rearrangements near the plasma membrane. Activation of Rac1, which is proposed to act upstream of the actin polymerization regulatory Wave 2 complex, has been found to correlate with enhanced macropinocytosis. One of ... More
Low-density Lipoprotein Receptor-related Protein-1 (LRP1) Mediates Autophagy and Apoptosis Caused by Helicobacter pylori VacA.
AuthorsYahiro K, Satoh M, Nakano M, Hisatsune J, Isomoto H, Sap J, Suzuki H, Nomura F, Noda M, Moss J, Hirayama T,
JournalJ Biol Chem
PubMed ID22822085
In Helicobacter pylori infection, vacuolating cytotoxin (VacA)-induced mitochondrial damage leading to apoptosis is believed to be a major cause of cell death. It has also been proposed that VacA-induced autophagy serves as a host mechanism to limit toxin-induced cellular damage. Apoptosis and autophagy are two dynamic and opposing processes that ... More
Large T antigen promotes JC virus replication in G2-arrested cells by inducing ATM- and ATR-mediated G2 checkpoint signaling.
AuthorsOrba Y, Suzuki T, Makino Y, Kubota K, Tanaka S, Kimura T, Sawa H,
JournalJ Biol Chem
PubMed ID19903823
Large T antigen (TAg) of the human polyomavirus JC virus (JCV) possesses DNA binding and helicase activities, which, together with various cellular proteins, are required for replication of the viral genome. We now show that JCV-infected cells expressing TAg accumulate in the G(2) phase of the cell cycle as a ... More
The utilization of pathogen-like cellular trafficking by single chain block copolymer.
Amphiphilic triblock copolymer, poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide), Pluronic P85, is unexpectedly shown to utilize sophisticated cellular trafficking mechanisms and enter brain microvessel endothelial cells and primary neurons that are poorly penetrable. Though caveolae serve as a primary entry site for the copolymer single chains, in cells devoid of caveolae, the ... More
Rosiglitazone treatment prevents mitochondrial dysfunction in mutant huntingtin-expressing cells: possible role of peroxisome proliferator-activated receptor-gamma (PPARgamma) in the pathogenesis of Huntington disease.
AuthorsQuintanilla RA, Jin YN, Fuenzalida K, Bronfman M, Johnson GV,
JournalJ Biol Chem
PubMed ID18640979
Peroxisome proliferator-activated receptor-gamma (PPARgamma) is a member of the PPAR family of transcription factors. Synthetic PPARgamma agonists are used as oral anti-hyperglycemic drugs for the treatment of non-insulin-dependent diabetes. However, emerging evidence indicates that PPARgamma activators can also prevent or attenuate neurodegeneration. Given these previous findings, the focus of this ... More
Nucleotide excision repair-induced H2A ubiquitination is dependent on MDC1 and RNF8 and reveals a universal DNA damage response.
Chromatin modifications are an important component of the of DNA damage response (DDR) network that safeguard genomic integrity. Recently, we demonstrated nucleotide excision repair (NER)-dependent histone H2A ubiquitination at sites of ultraviolet (UV)-induced DNA damage. In this study, we show a sustained H2A ubiquitination at damaged DNA, which requires dynamic ... More
STIM1 gates the store-operated calcium channel ORAI1 in vitro.
AuthorsZhou Y, Meraner P, Kwon HT, Machnes D, Oh-hora M, Zimmer J, Huang Y, Stura A, Rao A, Hogan PG,
JournalNat Struct Mol Biol
PubMed ID20037597
Store-operated Ca(2+) entry through the plasma membrane Ca(2+) release-activated Ca(2+) (CRAC) channel in mammalian T cells and mast cells depends on the sensor protein stromal interaction molecule 1 (STIM1) and the channel subunit ORAI1. To study STIM1-ORAI1 signaling in vitro, we have expressed human ORAI1 in a sec6-4 strain of ... More
Efficient polyethylenimine-mediated gene delivery proceeds via a caveolar pathway in HeLa cells.
AuthorsGabrielson NP, Pack DW,
JournalJ Control Release
PubMed ID19217921
Most in vivo gene therapies will require cell-specific targeting. Although vector targeting through ligand attachment has met with success in generating gene delivery particles that are capable of specific cellular interactions, little attention has been given to the possible effects of such ligands on subsequent intracellular processing. In this study, ... More
GPR55 is a cannabinoid receptor that increases intracellular calcium and inhibits M current.
AuthorsLauckner JE, Jensen JB, Chen HY, Lu HC, Hille B, Mackie K,
JournalProc Natl Acad Sci U S A
PubMed ID18263732
The CB(1) cannabinoid receptor mediates many of the psychoactive effects of Delta(9)THC, the principal active component of cannabis. However, ample evidence suggests that additional non-CB(1)/CB(2) receptors may contribute to the behavioral, vascular, and immunological actions of Delta(9)THC and endogenous cannabinoids. Here, we provide further evidence that GPR55, a G protein-coupled ... More
Regulation of calcium/calmodulin-dependent kinase IV by O-GlcNAc modification.
AuthorsDias WB, Cheung WD, Wang Z, Hart GW,
JournalJ Biol Chem
PubMed ID19506079
Similar to phosphorylation, GlcNAcylation (the addition of O-GlcNAc to Ser(Thr) residues on polypeptides) is an abundant, dynamic, and inducible post-translational modification. GlcNAcylated proteins are crucial in regulating virtually all cellular processes, including signaling, cell cycle, and transcription. Here we show that calcium/calmodulin-dependent kinase IV (CaMKIV) is highly GlcNAcylated in vivo. ... More
A fluorimetry-based ssYFP secretion assay to monitor vasopressin-induced exocytosis in LLC-PK1 cells expressing aquaporin-2.
AuthorsNunes P, Hasler U, McKee M, Lu HA, Bouley R, Brown D,
JournalAm J Physiol Cell Physiol
PubMed ID18799651
Vasopressin (VP)-induced exocytosis was dissected in native and aquaporin-2 (AQP2)-expressing renal LLC-PK(1) cells by a fluorimetric exocytosis assay based on soluble secreted yellow fluorescent protein (ssYFP). YFP was targeted to the secretory pathway by addition of an 18-amino acid signal peptide from hen egg white lysozyme. Immunofluorescence labeling, together with ... 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
Identification and Characterization of a Novel Gene, dapr, Involved in Skeletal Muscle Differentiation and Protein Kinase B Signaling.
AuthorsVirtanen C, Paris J, Takahashi M,
JournalJ Biol Chem
PubMed ID19028694
The phosphatidylinositol 3-kinase (PI3K) signaling pathway has been associated with a variety of cellular functions ranging from cell cycle regulation to tissue development. Although years of research have extensively characterized this signaling pathway, little is known as to how specific cellular events are coordinated by its activation. Here we demonstrate ... More
A role for huntington disease protein in dendritic RNA granules.
AuthorsSavas JN, Ma B, Deinhardt K, Culver BP, Restituito S, Wu L, Belasco JG, Chao MV, Tanese N,
JournalJ Biol Chem
PubMed ID20185826
Regulated transport and local translation of mRNA in neurons are critical for modulating synaptic strength, maintaining proper neural circuitry, and establishing long term memory. Neuronal RNA granules are ribonucleoprotein particles that serve to transport mRNA along microtubules and control local protein synthesis in response to synaptic activity. Studies suggest that ... More
Insoluble, speckled cytosolic distribution of retinoic acid receptor alpha protein as a marker of hepatic stellate cell activation in vitro.
AuthorsMezaki Y, Yamaguchi N, Yoshikawa K, Miura M, Imai K, Itoh H, Senoo H,
JournalJ Histochem Cytochem
PubMed ID19332432
Hepatic stellate cells (HSCs) are the major site of retinoid storage, and their activation is a key process in liver fibrogenesis. We have previously shown that expression of the retinoic acid receptor alpha (RARalpha) is upregulated in activated rat HSCs at a posttranscriptional level and that these RARalpha proteins showed ... More
Functional protein delivery into neurons using polymeric nanoparticles.
AuthorsHasadsri L, Kreuter J, Hattori H, Iwasaki T, George JM,
JournalJ Biol Chem
PubMed ID19129199
An efficient route for delivering specific proteins and peptides into neurons could greatly accelerate the development of therapies for various diseases, especially those involving intracellular defects such as Parkinson disease. Here we report the novel use of polybutylcyanoacrylate nanoparticles for delivery of intact, functional proteins into neurons and neuronal cell ... More
Protective role of endogenous gangliosides for lysosomal pathology in a cellular model of synucleinopathies.
AuthorsWei J, Fujita M, Nakai M, Waragai M, Sekigawa A, Sugama S, Takenouchi T, Masliah E, Hashimoto M,
JournalAm J Pathol
PubMed ID19349362
Gangliosides may be involved in the pathogenesis of Parkinson's disease and related disorders, although the precise mechanisms governing this involvement remain unknown. In this study, we determined whether changes in endogenous ganglioside levels affect lysosomal pathology in a cellular model of synucleinopathy. For this purpose, dementia with Lewy body-linked P123H ... More