Cyclin G2 associates with protein phosphatase 2A catalytic and regulatory B' subunits in active complexes and induces nuclear aberrations and a G1/S phase cell cycle arrest.
AuthorsBennin David A; Don Aruni S Arachchige; Brake Tiffany; McKenzie Jennifer L; Rosenbaum Heidi; Ortiz Linette; DePaoli-Roach Anna A; Horne Mary C;
JournalJ Biol Chem
PubMed ID11956189
Cyclin G2, together with cyclin G1 and cyclin I, defines a novel cyclin family expressed in terminally differentiated tissues including brain and muscle. Cyclin G2 expression is up-regulated as cells undergo cell cycle arrest or apoptosis in response to inhibitory stimuli independent of p53 (Horne, M., Donaldson, K., Goolsby, G., ... More
Functional overexpression of gamma-secretase reveals protease-independent trafficking functions and a critical role of lipids for protease activity.
AuthorsWrigley JD, Schurov I, Nunn EJ, Martin AC, Clarke EE, Ellis S, Bonnert TP, Shearman MS, Beher D
JournalJ Biol Chem
PubMed ID15613471
'Presenilins appear to form the active center of gamma-secretase but require the presence of the integral membrane proteins nicastrin, anterior pharynx defective 1, and presenilin enhancer 2 for catalytic function. We have simultaneously overexpressed all of these polypeptides, and we demonstrate functional assembly of the enzyme complex, a substantial increase ... More
Toward the generation of rod and cone photoreceptors from mouse, monkey and human embryonic stem cells.
AuthorsOsakada F, Ikeda H, Mandai M, Wataya T, Watanabe K, Yoshimura N, Akaike A, Akaike A, Sasai Y, Takahashi M,
JournalNat Biotechnol
PubMed ID18246062
'We previously reported the differentiation of mouse embryonic stem (ES) cells into retinal progenitors. However, these progenitors rarely differentiate into photoreceptors unless they are cultured with embryonic retinal tissues. Here we show the in vitro generation of putative rod and cone photoreceptors from mouse, monkey and human ES cells by ... More
Dicer inactivation leads to progressive functional and structural degeneration of the mouse retina.
AuthorsDamiani D, Alexander JJ, O'Rourke JR, McManus M, Jadhav AP, Cepko CL, Hauswirth WW, Harfe BD, Strettoi E,
JournalJ Neurosci
PubMed ID18463241
'MicroRNAs (miRNAs) are small, highly conserved molecules that have been shown to regulate the expression of genes by binding to specific target mRNAs. Dicer, an RNase III endonuclease, is essential for the production and function of mature miRNAs, and removal of Dicer has been shown to disrupt many developmental processes. ... More
Viral nanoparticles as tools for intravital vascular imaging.
AuthorsLewis JD, Destito G, Zijlstra A, Gonzalez MJ, Quigley JP, Manchester M, Stuhlmann H
JournalNat Med
PubMed ID16501571
'A significant impediment to the widespread use of noninvasive in vivo vascular imaging techniques is the current lack of suitable intravital imaging probes. We describe here a new strategy to use viral nanoparticles as a platform for the multivalent display of fluorescent dyes to image tissues deep inside living organisms. ... More
MBD3 and HDAC1, two components of the NuRD complex, are localized at Aurora-A-positive centrosomes in M phase.
AuthorsSakai H, Urano T, Ookata K, Kim MH, Hirai Y, Saito M, Nojima Y, Ishikawa F
JournalJ Biol Chem
PubMed ID12354758
'MBD3, a component of the histone deacetylase NuRD complex, contains the methyl-CpG-binding domain (MBD), yet does not possess appreciable mCpG-specific binding activity. The functional significance of MBD3 in the NuRD complex remains enigmatic, partly because of the limited availability of biochemical approaches, such as immunoprecipitation, to analyze MBD3. In this ... More
Nuclear localization of a non-caspase truncation product of atrophin-1, with an expanded polyglutamine repeat, increases cellular toxicity.
AuthorsNucifora FC, Ellerby LM, Wellington CL, Wood JD, Herring WJ, Sawa A, Hayden MR, Dawson VL, Dawson TM, Ross CA
JournalJ Biol Chem
PubMed ID12464607
'Dentatorubral and pallidoluysian atrophy (DRPLA) is an autosomal dominant neurodegenerative disorder similar to Huntington's disease, with clinical manifestations including chorea, incoordination, ataxia, and dementia. It is caused by an expansion of a CAG trinucleotide repeat encoding polyglutamine in the atrophin-1 gene. Both patients and DRPLA transgenic mice have nuclear accumulation ... More
Murine cytomegalovirus M78 protein, a G protein-coupled receptor homologue, is a constituent of the virion and facilitates accumulation of immediate-early viral mRNA.
AuthorsOliveira SA, Shenk TE
JournalProc Natl Acad Sci U S A
PubMed ID11248062
'The M78 protein of murine cytomegalovirus exhibits sequence features of a G protein-coupled receptor. It is synthesized with early kinetics, it becomes partially colocalized with Golgi markers, and it is incorporated into viral particles. We have constructed a viral substitution mutant, SMsubM78, which lacks most of the M78 ORF. The ... More
Disruption of nuclear lamin organization blocks the elongation phase of DNA replication.
AuthorsMoir RD, Spann TP, Herrmann H, Goldman RD
JournalJ Cell Biol
PubMed ID10851016
'The role of nuclear lamins in DNA replication is unclear. To address this, nuclei were assembled in Xenopus extracts containing AraC, a reversible inhibitor that blocks near the onset of the elongation phase of replication. Dominant-negative lamin mutants lacking their NH(2)-terminal domains were added to assembled nuclei to disrupt lamin ... More
A functional YNKI motif in the short cytoplasmic tail of varicella-zoster virus glycoprotein gH mediates clathrin-dependent and antibody-independent endocytosis.
AuthorsPasieka TJ, Maresova L, Grose C,
JournalJ Virol
PubMed ID12634377
'The trafficking of varicella-zoster virus (VZV) gH was investigated under both infection and transfection conditions. In initial endocytosis assays performed in infected cells, the three glycoproteins gE, gI, and gB served as positive controls for internalization from the plasma membrane. Subsequently, we discovered that gH in VZV-infected cells was also ... More
Cohesin defects lead to premature sister chromatid separation, kinetochore dysfunction, and spindle-assembly checkpoint activation.
AuthorsHoque MT, Ishikawa F
JournalJ Biol Chem
PubMed ID12200439
'Scc1/Mcd1 is a component of the cohesin complex that plays an essential role in sister chromatid cohesion in eukaryote cells. Knockout experiments of this gene have been described in budding yeast, fission yeast, and chicken cells, but no study has been reported on human Scc1 thus far. In this study, ... More
A functional angiotensin II receptor-GFP fusion protein: evidence for agonist-dependent nuclear translocation.
AuthorsChen R, Mukhin YV, Garnovskaya MN, Thielen TE, Iijima Y, Huang C, Raymond JR, Ullian ME, Paul RV
JournalAm J Physiol Renal Physiol
PubMed ID10966923
'We constructed an expression vector for a fusion protein [ANG II type 1a receptor-green fluorescent protein (AT(1a)R-GFP)] consisting of enhanced GFP attached to the COOH terminus of the rat AT(1a)R. Chinese hamster ovary (CHO) cells transfected with AT(1a)R-GFP demonstrated specific, high-affinity (125)I-labeled ANG II binding (IC(50) 21 nM). ANG II ... More
Diverse microglial motility behaviors during clearance of dead cells in hippocampal slices.
AuthorsPetersen MA, Dailey ME
JournalGlia
PubMed ID15042586
'We used two-channel three-dimensional time-lapse fluorescence confocal imaging in live rat hippocampal slice cultures (1-7 days in vitro) to determine the motility behaviors of activated microglia as they engage dead and dying cells following traumatic brain tissue injury. Live microglia were labeled with a fluorescently conjugated lectin (IB(4)), and dead ... More
Kinesin Superfamily Motor Protein KIF17 and mLin-10 in NMDA Receptor-Containing Vesicle Transport
'Experiments with vesicles containing N-methyl-D-aspartate (NMDA)receptor 2B (NR2B subunit) show that they are transported alongmicrotubules by KIF17, a neuron-specific molecular motor in neuronaldendrites. Selective transport is accomplished by direct interaction of theKIF17 tail with a PDZ domain of mLin-10 (Mint1/X11), which is aconstituent of a large protein complex including mLin-2 ... More
Visible diode lasers can be used for flow cytometric immunofluorescence and DNA analysis.
AuthorsDoornbos RM, De Grooth BG, Kraan YM, Van Der Poel CJ, Greve J
JournalCytometry
PubMed ID8187585
'This report describes a feasibility study concerning the use of a visible diode laser for two important fluorescence applications in a flow cytometer. With a 3 mW 635 nm diode laser, we performed immunofluorescence measurements using the fluorophore allophycocyanin (APC). We have measured CD8 positive lymphocytes with a two-step labeling ... More
Dynamics of transitional endoplasmic reticulum sites in vertebrate cells.
AuthorsHammond AT, Glick BS
JournalMol Biol Cell
PubMed ID10982397
'A typical vertebrate cell contains several hundred sites of transitional ER (tER). Presumably, tER sites generate elements of the ER-Golgi intermediate compartment (ERGIC), and ERGIC elements then generate Golgi cisternae. Therefore, characterizing the mechanisms that influence tER distribution may shed light on the dynamic behavior of the Golgi. We explored ... More
Zebrafish vasa RNA but not its protein is a component of the germ plasm and segregates asymmetrically before germline specification.
AuthorsKnaut H, Pelegri F, Bohmann K, Schwarz H, Nüsslein-Volhard C
JournalJ Cell Biol
PubMed ID10811828
'Work in different organisms revealed that the vasa gene product is essential for germline specification. Here, we describe the asymmetric segregation of zebrafish vasa RNA, which distinguishes germ cell precursors from somatic cells in cleavage stage embryos. At the late blastula (sphere) stage, vasa mRNA segregation changes from asymmetric to ... More
Targeting Janus kinase 3 in mast cells prevents immediate hypersensitivity reactions and anaphylaxis.
AuthorsMalaviya R, Zhu D, Dibirdik I, Uckun FM
JournalJ Biol Chem
PubMed ID10480916
'Janus kinase 3 (JAK3), a member of the Janus family protein-tyrosine kinases, is expressed in mast cells, and its enzymatic activity is enhanced by IgE receptor/FcepsilonRI cross-linking. Selective inhibition of JAK3 in mast cells with 4-(4''-hydroxylphenyl)-amino-6, 7-dimethoxyquinazoline) (WHI-P131) blocked the phospholipase C activation, calcium mobilization, and activation of microtubule-associated protein ... More
Applications of cytotoxicity assays and pre-lethal mechanistic assays for assessment of human hepatotoxicity potential.
AuthorsXu JJ, Diaz D, O'Brien PJ
JournalChem Biol Interact
PubMed ID15522265
'While drug toxicity (especially hepatotoxicity) is the most frequent reason cited for withdrawal of an approved drug, no simple solution exists to adequately predict such adverse events. Simple cytotoxicity assays in HepG2 cells are relatively insensitive to human hepatotoxic drugs in a retrospective analysis of marketed pharmaceuticals. In comparison, a ... More
Counting single molecules in sub-nanolitre droplets.
AuthorsRane TD, Puleo CM, Liu KJ, Zhang Y, Lee AP, Wang TH,
JournalLab Chip
PubMed ID20066242
'We demonstrate single biomolecule detection and quantification within sub-nanolitre droplets through application of Cylindrical Illumination Confocal Spectroscopy (CICS) and droplet confinement within a retractable microfluidic constriction.' ... More
DNA labeling in living cells.
AuthorsMartin RM, Leonhardt H, Cardoso MC
JournalCytometry A
PubMed ID16082711
'BACKGROUND: Live cell fluorescence microscopy experiments often require visualization of the nucleus and the chromatin to determine the nuclear morphology or the localization of nuclear compartments. METHODS: We compared five different DNA dyes, TOPRO-3, TOTO-3, propidium iodide, Hoechst 33258, and DRAQ5, to test their usefulness in live cell experiments with ... More
Fibroblasts as host cells in latent leishmaniosis.
AuthorsBogdan C, Donhauser N, Döring R, Röllinghoff M, Diefenbach A, Rittig MG
JournalJ Exp Med
PubMed ID10859337
'Intracellular parasites are known to persist lifelong in mammalian hosts after the clinical cure of the disease, but the mechanisms of persistence are poorly understood. Here, we show by confocal laser microscopy that in the draining lymph nodes of mice that had healed a cutaneous infection with Leishmania major, 40% ... More
The organization of replication and transcription.
AuthorsCook PR
JournalScience
PubMed ID10364545
'Models for replication and transcription often display polymerases that track like locomotives along their DNA templates. However, recent evidence supports an alternative model in which DNA and RNA polymerases are immobilized by attachment to larger structures, where they reel in their templates and extrude newly made nucleic acids. These polymerases ... More
Cyclin A is destroyed in prometaphase and can delay chromosome alignment and anaphase.
Authorsden Elzen N, Pines J
JournalJ Cell Biol
PubMed ID11285279
'Mitosis is controlled by the specific and timely degradation of key regulatory proteins, notably the mitotic cyclins that bind and activate the cyclin-dependent kinases (Cdks). In animal cells, cyclin A is always degraded before cyclin B, but the exact timing and the mechanism underlying this are not known. Here we ... More
FRET studies of the interaction of dimeric cyanine dyes with DNA.
AuthorsLaib S, Seeger S
JournalJ Fluoresc
PubMed ID15615044
'Fluorescence Resonance Energy Transfer (FRET) is a powerful tool to determine distances between chromophores bound to macromolecules, since the efficiency of the energy transfer from an initially excited donor to an acceptor strongly depends on the distance between the two dye molecules. The structure of the noncovalent complex of double-strand ... More
Methods and applications of three-color confocal imaging.
AuthorsPaddock SW, Hazen EJ, De Vries PJ
JournalBiotechniques
PubMed ID8994659
A simple method for constructing two- and three-color merged images from grayscale confocal fluorescence images using Adobe Photoshop is outlined. Various computer methods for manipulating and displaying the images are discussed in light of several recent biomedical applications of multi-label confocal microscopy. ... More
Approaches for monitoring nuclear translation.
AuthorsIborra FJ, Jackson DA, Cook PR
JournalMethods Mol Biol
PubMed ID14770000
The nuclear membrane is the defining feature of eukaryotes. It divides the cell into two functionally specialized compartments, and it is widely assumed that translation is restricted to only one: the cytoplasm. However, recent results suggest that some translation takes place in nuclei closely coupled to transcription. Various labeling techniques ... More
Evidence for stroke-induced neurogenesis in the human brain.
AuthorsJin K, Wang X, Xie L, Mao XO, Zhu W, Wang Y, Shen J, Mao Y, Banwait S, Greenberg DA
JournalProc Natl Acad Sci U S A
PubMed ID16924107
Experimental stroke in rodents stimulates neurogenesis and migration of newborn neurons from their sites of origin into ischemic brain regions. We report that in patients with stroke, cells that express markers associated with newborn neurons are present in the ischemic penumbra surrounding cerebral cortical infarcts, where these cells are preferentially ... More
Math1: an essential gene for the generation of inner ear hair cells.
AuthorsBermingham NA, Hassan BA, Price SD, Vollrath MA, Ben-Arie N, Eatock RA, Bellen HJ, Lysakowski A, Zoghbi HY
JournalScience
PubMed ID10364557
The mammalian inner ear contains the cochlea and vestibular organs, which are responsible for hearing and balance, respectively. The epithelia of these sensory organs contain hair cells that function as mechanoreceptors to transduce sound and head motion. The molecular mechanisms underlying hair cell development and differentiation are poorly understood. Math1, ... More
An olfactory sensory map in the fly brain.
AuthorsVosshall LB, Wong AM, Axel R
JournalCell
PubMed ID10943836
We have isolated the "complete" repertoire of genes encoding the odorant receptors in Drosophila and employ these genes to provide a molecular description of the organization of the peripheral olfactory system. The repertoire of Drosophila odorant receptors is encoded by 57 genes. Individual sensory neurons are likely to express only ... More
A chemosensory gene family encoding candidate gustatory and olfactory receptors in Drosophila.
AuthorsScott K, Brady R, Cravchik A, Morozov P, Rzhetsky A, Zuker C, Axel R
JournalCell
PubMed ID11257221
A novel family of candidate gustatory receptors (GRs) was recently identified in searches of the Drosophila genome. We have performed in situ hybridization and transgene experiments that reveal expression of these genes in both gustatory and olfactory neurons in adult flies and larvae. This gene family is likely to encode ... More
Hrs regulates endosome membrane invagination and tyrosine kinase receptor signaling in Drosophila.
Signaling through tyrosine kinase receptors (TKRs) is thought to be modulated by receptor-mediated endocytosis and degradation of the receptor in the lysosome. However, factors that regulate endosomal sorting of TKRs are largely unknown. Here, we demonstrate that Hrs (Hepatocyte growth factor-regulated tyrosine kinase substrate) is one such factor. Electron microscopy ... More
A spatial map of olfactory receptor expression in the Drosophila antenna.
AuthorsVosshall LB, Amrein H, Morozov PS, Rzhetsky A, Axel R
JournalCell
PubMed ID10089887
Insects provide an attractive system for the study of olfactory sensory perception. We have identified a novel family of seven transmembrane domain proteins, encoded by 100 to 200 genes, that is likely to represent the family of Drosophila odorant receptors. Members of this gene family are expressed in topographically defined ... More
Proprioceptor pathway development is dependent on Math1.
AuthorsBermingham NA, Hassan BA, Wang VY, Fernandez M, Banfi S, Bellen HJ, Fritzsch B, Zoghbi HY
JournalNeuron
PubMed ID11395003
The proprioceptive system provides continuous positional information on the limbs and body to the thalamus, cortex, pontine nucleus, and cerebellum. We showed previously that the basic helix-loop-helix transcription factor Math1 is essential for the development of certain components of the proprioceptive pathway, including inner-ear hair cells, cerebellar granule neurons, and ... More
An ent-kaurene that inhibits mitotic chromosome movement and binds the kinetochore protein ran-binding protein 2.
AuthorsRundle NT, Nelson J, Flory MR, Joseph J, Th'ng J, Aebersold R, Dasso M, Andersen RJ, Roberge M
JournalACS Chem Biol
PubMed ID17168522
Using a chemical genetics screen, we have identified ent-15-oxokaurenoic acid (EKA) as a chemical that causes prolonged mitotic arrest at a stage resembling prometaphase. EKA inhibits the association of the mitotic motor protein centromeric protein E with kinetochores and inhibits chromosome movement. Unlike most antimitotic agents, EKA does not inhibit ... More
Optimal processing method to obtain four-color confocal fluorescent images of the cytoskeleton and nucleus in three-dimensional chondrocyte cultures.
AuthorsBlanc A, Tran-Khanh N, Filion D, Buschmann MD
JournalJ Histochem Cytochem
PubMed ID15933071
Tissue engineering of articular cartilage requires accurate imaging of the chondrocyte cytoskeleton. Past studies have applied various fixation and permeabilization protocols without optimization of parameters. In this study, we have examined procedures using glutaraldehyde and paraformaldehyde as fixatives and Triton X-100 and Octyl-POE as permeabilizing detergents. A four-color fluorescence confocal ... More
Interference by huntingtin and atrophin-1 with cbp-mediated transcription leading to cellular toxicity.
AuthorsNucifora FC, Sasaki M, Peters MF, Huang H, Cooper JK, Yamada M, Takahashi H, Tsuji S, Troncoso J, Dawson VL, Dawson TM, Ross CA
JournalScience
PubMed ID11264541
Expanded polyglutamine repeats have been proposed to cause neuronal degeneration in Huntington's disease (HD) and related disorders, through abnormal interactions with other proteins containing short polyglutamine tracts such as the transcriptional coactivator CREB binding protein, CBP. We found that CBP was depleted from its normal nuclear location and was present ... More
Lymphocyte apoptosis induced by CD95 (APO-1/Fas) ligand-expressing tumor cells--a mechanism of immune evasion?
AuthorsStrand S, Hofmann WJ, Hug H, Müller M, Otto G, Strand D, Mariani SM, Stremmel W, Krammer PH, Galle PR
JournalNat Med
PubMed ID8946836
The CD95 (APO-1/Fas) system is an important mediator of T-cell cytotoxicity. We investigated this system in 22 hepatocellular carcinomas (HCCs) from patients. All HCCs had partially or completely lost the expression of the CD95 receptor constitutively expressed by normal liver cells and might thus evade CD95-mediated killing. We also considered ... More
Chemical transformations in individual ultrasmall biomimetic containers.
AuthorsChiu DT, Wilson CF, Ryttsén F, Strömberg A, Farre C, Karlsson A, Nordholm S, Gaggar A, Modi BP, Moscho A, Garza-López RA, Orwar O, Zare RN
JournalScience
PubMed ID10082457
Individual phospholipid vesicles, 1 to 5 micrometers in diameter, containing a single reagent or a complete reaction system, were immobilized with an infrared laser optical trap or by adhesion to modified borosilicate glass surfaces. Chemical transformations were initiated either by electroporation or by electrofusion, in each case through application of ... More
DNA ligase IV mutations identified in patients exhibiting developmental delay and immunodeficiency.
AuthorsO'Driscoll M, Cerosaletti KM, Girard PM, Dai Y, Stumm M, Kysela B, Hirsch B, Gennery A, Palmer SE, Seidel J, Gatti RA, Varon R, Oettinger MA, Neitzel H, Jeggo PA, Concannon P
JournalMol Cell
PubMed ID11779494
DNA ligase IV functions in DNA nonhomologous end-joining and V(D)J recombination. Four patients with features including immunodeficiency and developmental and growth delay were found to have mutations in the gene encoding DNA ligase IV (LIG4). Their clinical phenotype closely resembles the DNA damage response disorder, Nijmegen breakage syndrome (NBS). Some ... More
DNA staining for fluorescence and laser confocal microscopy.
AuthorsSuzuki T, Fujikura K, Higashiyama T, Takata K
JournalJ Histochem Cytochem
PubMed ID9010468
We examined five nucleic acid binding fluorescent dyes, propidium iodide, SYBR Green I, YO-PRO-1, TOTO-3, and TO-PRO-3, for nuclear DNA staining, visualized by fluorescence and laser confocal microscopy. The optimal concentration, co-staining of RNA, and bleaching speeds were examined. SYBR Green I and TO-PRO-3 almost preferentially stained the nuclear DNA, ... More
Role for centromeric heterochromatin and PML nuclear bodies in the cellular response to foreign DNA.
AuthorsBishop CL, Ramalho M, Nadkarni N, May Kong W, Higgins CF, Krauzewicz N,
JournalMol Cell Biol
PubMed ID16537904
Nuclear spatial positioning plays an important role in the epigenetic regulation of eukaryotic gene expression. Here we show a role for nuclear spatial positioning in regulating episomal transgenes that are delivered by virus-like particles (VLPs). VLPs mediate the delivery of plasmid DNA (pDNA) to cell nuclei but lack viral factors ... More
Glutathione depletion is necessary for apoptosis in lymphoid cells independent of reactive oxygen species formation.
AuthorsFranco R, Panayiotidis MI, Cidlowski JA,
JournalJ Biol Chem
PubMed ID17724027
Changes in the intracellular redox environment of cells have been reported to be critical for the activation of apoptotic enzymes and the progression of programmed cell death. Glutathione (GSH) depletion is an early hallmark observed in apoptosis, and we have demonstrated that GSH efflux during death receptor-mediated apoptosis occurs via ... More
High concordance of drug-induced human hepatotoxicity with in vitro cytotoxicity measured in a novel cell-based model using high content screening.
AuthorsO'Brien PJ, Irwin W, Diaz D, Howard-Cofield E, Krejsa CM, Slaughter MR, Gao B, Kaludercic N, Angeline A, Bernardi P, Brain P, Hougham C,
JournalArch Toxicol
PubMed ID16598496
To develop and validate a practical, in vitro, cell-based model to assess human hepatotoxicity potential of drugs, we used the new technology of high content screening (HCS) and a novel combination of critical model features, including (1) use of live, human hepatocytes with drug metabolism capability, (2) preincubation of cells ... More
Mechanism of Ca2+ disruption in Alzheimer's disease by presenilin regulation of InsP(3) receptor channel gating.
AuthorsCheung KH, Shineman D, Müller M, Cárdenas C, Mei L, Yang J, Tomita T, Iwatsubo T, Lee VM, Foskett JK,
JournalNeuron
PubMed ID18579078
Mutations in presenilins (PS) are the major cause of familial Alzheimer's disease (FAD) and have been associated with calcium (Ca2+) signaling abnormalities. Here, we demonstrate that FAD mutant PS1 (M146L)and PS2 (N141I) interact with the inositol 1,4,5-trisphosphate receptor (InsP3R) Ca2+ release channel and exert profound stimulatory effects on its gating ... More
Doublecortin is a microtubule-associated protein and is expressed widely by migrating neurons.
AuthorsGleeson JG, Lin PT, Flanagan LA, Walsh CA
JournalNeuron
PubMed ID10399933
Doublecortin (DCX) is required for normal migration of neurons into the cerebral cortex, since mutations in the human gene cause a disruption of cortical neuronal migration. To date, little is known about the distribution of DCX protein or its function. Here, we demonstrate that DCX is expressed in migrating neurons ... More
CPEB, maskin, and cyclin B1 mRNA at the mitotic apparatus: implications for local translational control of cell division.
AuthorsGroisman I, Huang YS, Mendez R, Cao Q, Theurkauf W, Richter JD
JournalCell
PubMed ID11081630
In Xenopus development, the expression of several maternal mRNAs is regulated by cytoplasmic polyadenylation. CPEB and maskin, two factors that control polyadenylation-induced translation are present on the mitotic apparatus of animal pole blastomeres in embryos. Cyclin B1 protein and mRNA, whose translation is regulated by polyadenylation, are colocalized with CPEB ... More
The localization of human cyclins B1 and B2 determines CDK1 substrate specificity and neither enzyme requires MEK to disassemble the Golgi apparatus.
AuthorsDraviam VM, Orrechia S, Lowe M, Pardi R, Pines J
JournalJ Cell Biol
PubMed ID11238451
In this paper, we show that substrate specificity is primarily conferred on human mitotic cyclin-dependent kinases (CDKs) by their subcellular localization. The difference in localization of the B-type cyclin-CDKs underlies the ability of cyclin B1-CDK1 to cause chromosome condensation, reorganization of the microtubules, and disassembly of the nuclear lamina and ... More
Drosophila Aurora A kinase is required to localize D-TACC to centrosomes and to regulate astral microtubules.
Disruption of the function of the A-type Aurora kinase of Drosophila by mutation or RNAi leads to a reduction in the length of astral microtubules in syncytial embryos, larval neuroblasts, and cultured S2 cells. In neuroblasts, it can also lead to loss of an organized centrosome and its associated aster ... More
Nucleus labeling or membrane labeling for studying the proliferation of drug treated cells?
AuthorsBoutonnat J, Barbier M, Ronot X, Seigneurin D
JournalMorphologie
PubMed ID11048293
Proliferation and multidrug resistance status are key predictors of therapeutic outcome in acute myeloid leukemia (AML). This study compared cell cycle analysis (nuclear labeling) with cell division analysis (membrane labeling, PKH67) for studying the proliferation of cells cultured with daunorubicin (DNR) and/or Cytarabine (Ara-C), drugs commonly used in AML treatment. ... More
Transport of nucleosome core particles in semidilute DNA solutions.
AuthorsMangenot S, Keller S, Rädler J
JournalBiophys J
PubMed ID12944295
We studied the diffusion of native and trypsinized nucleosome core particles (NCPs), in aqueous solution and in concentrated DNA solutions (0.25-100 mg/ml) using fluorescence correlation spectroscopy (FCS). The highest DNA concentrations studied mimic the DNA density inside the cell nucleus. The diffusion coefficient of freely diffusing NCPs depends on the ... More
A role for nuclear lamins in nuclear envelope assembly.
The molecular interactions responsible for nuclear envelope assembly after mitosis are not well understood. In this study, we demonstrate that a peptide consisting of the COOH-terminal domain of Xenopus lamin B3 (LB3T) prevents nuclear envelope assembly in Xenopus interphase extracts. Specifically, LB3T inhibits chromatin decondensation and blocks the formation of ... More
Multipurpose high sensitivity luminescence analyzer (LUANA): use in gel electrophoresis.
AuthorsNeri D, Prospero T, Petrul H, Winter G, Browne M, Vanderpant L
JournalBiotechniques
PubMed ID8800693
Many applications in molecular biology require the rapid and high-sensitivity detection of biological macromolecules. Here we describe a luminescence analyzer (LUANA), featuring xenon lamp-based illumination and a cooled CCD camera as detector. Luminescent samples (gels, membranes or microplates) are placed in a light-tight chamber, and computer software is used to ... More
Simultaneous imaging and functional assessment of cytoskeletal protein connections in passively loaded single muscle cells.
AuthorsShah SB, Lieber RL
JournalJ Histochem Cytochem
PubMed ID12502751
We describe a novel system that permits simultaneous confocal imaging of protein interactions and measurement of cell mechanical properties during passive loading. A mechanical apparatus was designed to replace the stage of a confocal microscope, enabling cell manipulation, force transduction, and imaging. In addition, image processing algorithms were developed to ... More
The anti-inflammatory sesquiterpene lactone helenalin inhibits the transcription factor NF-kappaB by directly targeting p65.
AuthorsLyss G, Knorre A, Schmidt TJ, Pahl HL, Merfort I
JournalJ Biol Chem
PubMed ID9837931
The sesquiterpene lactone helenalin is a potent anti-inflammatory drug whose molecular mechanism of action remains unclear despite numerous investigations. We have previously shown that helenalin and other sesquiterpene lactones selectively inhibit activation of the transcription factor NF-kappaB, a central mediator of the human immune response. These drugs must target a ... More
In vivo analysis of human multidrug resistance protein 1 (MRP1) activity using transient expression of fluorescently tagged MRP1.
AuthorsRajagopal A, Pant AC, Simon SM, Chen Y
JournalCancer Res
PubMed ID11809686
The multidrug resistance protein 1 (MRP1) contributes cellular resistance to a wide array of physiological toxins and chemotherapeutic agents. Its in vivo activity has been studied primarily in cells that have been continuously drug selected, culture conditions that might confound the effects of MRP1 expression with the effects of a ... More
The positioning and dynamics of origins of replication in the budding yeast nucleus.
AuthorsHeun P, Laroche T, Raghuraman MK, Gasser SM
JournalJ Cell Biol
PubMed ID11266454
We have analyzed the subnuclear position of early- and late-firing origins of DNA replication in intact yeast cells using fluorescence in situ hybridization and green fluorescent protein (GFP)-tagged chromosomal domains. In both cases, origin position was determined with respect to the nuclear envelope, as identified by nuclear pore staining or ... More
Chaperones Hsp70 and Hsp40 suppress aggregate formation and apoptosis in cultured neuronal cells expressing truncated androgen receptor protein with expanded polyglutamine tract.
AuthorsKobayashi Y, Kume A, Li M, Doyu M, Hata M, Ohtsuka K, Sobue G
JournalJ Biol Chem
PubMed ID10722721
Spinal and bulbar muscular atrophy (SBMA) is one of a group of human inherited neurodegenerative diseases caused by polyglutamine expansion. We have previously demonstrated that the SBMA gene product, the androgen receptor protein, is toxic and aggregates when truncated. Heat shock proteins function as molecular chaperones, which recognize and renaturate ... More
Stable correction of a genetic deficiency in human cells by an episome carrying a 115 kb genomic transgene.
AuthorsWade-Martins R, White RE, Kimura H, Cook PR, James MR
JournalNat Biotechnol
PubMed ID11101814
Persistent expression of a transgene at therapeutic levels is required for successful gene therapy, but many small vectors with heterologous promoters are prone to vector loss and transcriptional silencing. The delivery of genomic DNA would enable genes to be transferred as complete loci, including regulatory sequences, introns, and native promoter ... More
Scurfin (FOXP3) acts as a repressor of transcription and regulates T cell activation.
AuthorsSchubert LA, Jeffery E, Zhang Y, Ramsdell F, Ziegler SF
JournalJ Biol Chem
PubMed ID11483607
We have recently identified and cloned Foxp3, the gene defective in mice with the scurfy mutation. The immune dysregulation documented in these mice and in humans with mutations in the orthologous gene indicates that the foxp3 gene product, scurfin, is involved in the regulation of T cell activation and differentiation. ... More
Functional cardiac cell constructs on cellulose-based scaffolding.
AuthorsEntcheva E, Bien H, Yin L, Chung CY, Farrell M, Kostov Y
JournalBiomaterials
PubMed ID15147821
Cellulose and its derivatives have been successfully employed as biomaterials in various applications, including dialysis membranes, diffusion-limiting membranes in biosensors, in vitro hollow fibers perfusion systems, surfaces for cell expansion, etc. In this study, we tested the potential of cellulose acetate (CA) and regenerated cellulose (RC) scaffolds for growing functional ... More
Lamin A/C expression is a marker of mouse and human embryonic stem cell differentiation.
AuthorsConstantinescu D, Gray HL, Sammak PJ, Schatten GP, Csoka AB
JournalStem Cells
PubMed ID16179429
Nuclear lamins comprise the nuclear lamina, a scaffold-like structure that lines the inner nuclear membrane. B-type lamins are present in almost all cell types, but A-type lamins are expressed predominantly in differentiated cells, suggesting a role in maintenance of the differentiated state. Previous studies have shown that lamin A/C is ... More
Interferon-alpha induces nmi-IFP35 heterodimeric complex formation that is affected by the phosphorylation of IFP35.
Nmi and IFP35 are interferon (IFN)-induced proteins. In cells treated with IFN-gamma, Nmi enhances the association of transcription co-activator CBP/p300 with signal transducer and activator of transcription proteins, and IFP35 forms a high molecular weight cytosolic complex of unknown constituents. Here we show that Nmi and IFP35 co-immunoprecipitate with an ... More
The RACK1 homologue from Trypanosoma brucei is required for the onset and progression of cytokinesis.
AuthorsRothberg KG, Burdette DL, Pfannstiel J, Jetton N, Singh R, Ruben L
JournalJ Biol Chem
PubMed ID16469736
The receptor for activated C kinase 1 (RACK1) is a conserved scaffold protein that helps regulate a range of cell activities including cell growth, shape, and protein translation. We report that a homologue of RACK1 is required for cytokinesis in pathogenic Trypanosoma brucei. The protein, referred to as TRACK, is ... More
On the possibility of long-wavelength long-lifetime high-quantum-yield luminophores.
AuthorsLakowicz JR, Piszczek G, Kang JS
JournalAnal Biochem
PubMed ID11141307
We describe an approach to creating a new class of luminophores which display both long wavelength emissions exceeding 600 nm and long lifetimes. These luminophores are based on resonance energy transfer (RET) from a long lifetime donor to a short lifetime but long wavelength acceptor. We demonstrated the possibility of ... More
Replicon clusters are stable units of chromosome structure: evidence that nuclear organization contributes to the efficient activation and propagation of S phase in human cells.
AuthorsJackson DA, Pombo A
JournalJ Cell Biol
PubMed ID9508763
In proliferating cells, DNA synthesis must be performed with extreme precision. We show that groups of replicons, labeled together as replicon clusters, form stable units of chromosome structure. HeLa cells were labeled with 5-bromodeoxyuridine (BrdU) at different times of S phase. At the onset of S phase, clusters of replicons ... More
Olfactory responses in a gustatory organ of the malaria vector mosquito Anopheles gambiae.
AuthorsKwon HW, Lu T, Rützler M, Zwiebel LJ
JournalProc Natl Acad Sci U S A
PubMed ID16938890
The proboscis is an important head appendage in insects that has primarily been thought to process gustatory information during food intake. Indeed, in Drosophila and other insects in which they have been identified, most gustatory receptors are expressed in proboscis neurons. Our previous characterization of the expression of AgOR7, a ... More
Homogeneous Cell- and Bead-Based Assays for High Throughput Screening Using Fluorometric Microvolume Assay Technology.
AuthorsMiraglia S, Swartzman EE, Mellentin-Michelotti J, Evangelista L, Smith C, Gunawan I, Lohman K, Goldberg EM, Manian B, Yuan PM
JournalJ Biomol Screen
PubMed ID10838439
High throughput drug screening has become a critical component of the drug discovery process. The screening of libraries containing hundreds of thousands of compounds has resulted in a requirement for assays and instrumentation that are amenable to nonradioactive formats and that can be miniaturized. Homogeneous assays that minimize upstream automation ... More
Drosophila aurora B kinase is required for histone H3 phosphorylation and condensin recruitment during chromosome condensation and to organize the central spindle during cytokinesis.
AuthorsGiet R, Glover DM
JournalJ Cell Biol
PubMed ID11266459
Aurora/Ipl1-related kinases are a conserved family of enzymes that have multiple functions during mitotic progression. Although it has been possible to use conventional genetic analysis to dissect the function of aurora, the founding family member in Drosophila (Glover, D.M., M.H. Leibowitz, D.A. McLean, and H. Parry. 1995. Cell. 81:95-105), the ... More
Sensitive and reliable JC-1 and TOTO-3 double staining to assess mitochondrial transmembrane potential and plasma membrane integrity: interest for cell death investigations.
AuthorsZuliani T, Duval R, Jayat C, Schnébert S, André P, Dumas M, Ratinaud MH
JournalCytometry A
PubMed ID12879456
BACKGROUND: Apoptosis is currently studied by flow cytometry with mitochondrial membrane potential (Deltapsimt) and membrane integrity fluorochromes. Rhodamine 123 and DiOC6(3) remain controversial to identify cells displaying a low Deltapsimt. JC-1 constitutes a good Deltapsimt indicator, due to a fluorescence shift from green to orange emission, according to the increase ... More
Characteristics of a novel deep red/infrared fluorescent cell-permeant DNA probe, DRAQ5, in intact human cells analyzed by flow cytometry, confocal and multiphoton microscopy.
AuthorsSmith PJ, Blunt N, Wiltshire M, Hoy T, Teesdale-Spittle P, Craven MR, Watson JV, Amos WB, Errington RJ, Patterson LH
JournalCytometry
PubMed ID10918279
BACKGROUND: The multiparameter fluorometric analysis of intact and fixed cells often requires the use of a nuclear DNA discrimination signal with spectral separation from visible range fluorochromes. We have developed a novel deep red fluorescing bisalkylaminoanthraquinone, DRAQ5 (Ex(lambdamax) 646 nm; Em(lambdamax) 681 nm; Em(lambdarange) 665->800 nm), with high affinity for ... More
Solution structure of a DNA complex with the fluorescent bis-intercalator TOTO determined by NMR spectroscopy.
AuthorsSpielmann HP, Wemmer DE, Jacobsen JP
JournalBiochemistry
PubMed ID7612596
We have used two-dimensional 1H NMR spectroscopy to determine the solution structure of the DNA oligonucleotide d(5'-CGCTAGCG-3')2 complexed with the bis-intercalating dye 1,1'-(4,4,8,8-tetramethyl-4,8-diazaundecamethylene)bis[4-(3-methyl -2,3- dihydrobenzo-1,3-thiazolyl-2-methylidene)qui nolinium] tetraiodide (TOTO). The determination of the structure was based on total relaxation matrix analysis of the NOESY cross-peak intensities using the program MARDIGRAS. Improved ... More
Regional loss of the mitochondrial membrane potential in the hepatocyte is rapidly followed by externalization of phosphatidylserines at that specific site during apoptosis.
AuthorsBlom WM, de Bont HJ, Nagelkerke JF
JournalJ Biol Chem
PubMed ID12538597
The spatio-temporal relationship between a decrease in the mitochondrial membrane potential (MMP) and externalization of phosphatidylserines (PS) during induction of apoptosis was investigated in single freshly isolated hepatocytes. Apoptosis was induced in the hepatocytes in three different ways: attack by activated Natural Killer cells, exposure to ATP, or exposure to ... More
Effects of hyperthyroidism in the development of the appendicular skeleton and muscles of zebrafish, with notes on evolutionary developmental pathology (Evo-Devo-Path).
AuthorsShkil F, Siomava N, Voronezhskaya E, Diogo R
JournalSci Rep
PubMed ID30931985
'The hypothalamus-pituitary-thyroid (HPT) axis plays a crucial role in the metabolism, homeostasis, somatic growth and development of teleostean fishes. Thyroid hormones regulate essential biological functions such as growth and development, regulation of stress, energy expenditure, tissue compound, and psychological processes. Teleost thyroid follicles produce the same thyroid hormones as in ... More
NF-?B p65 directs sex-specific neuroprotection in human neurons.
'Protection of neurons against oxidative stress is crucial during neuronal development, maintenance and for treating neurodegenerative diseases. However, little is known about the molecular mechanisms underlying sex-specific maturation and survival of neurons. In the present study, we demonstrate NF-?B-p65 mediated neuroprotection in human glutamatergic neurons differentiated from inferior turbinate stem ... More
The genomics of ecological flexibility, large brains, and long lives in capuchin monkeys revealed with fecalFACS.
Authors
JournalProc Natl Acad Sci U S A
PubMed ID33574059
Systematic functional analysis of rab GTPases reveals limits of neuronal robustness to environmental challenges in flies.
Authors
JournalElife
PubMed ID33666175
Human RECQ1 promotes restart of replication forks reversed by DNA topoisomerase I inhibition.
Authors
JournalNat Struct Mol Biol
PubMed ID23396353
Deletion of Mecp2 in Sim1-expressing neurons reveals a critical role for MeCP2 in feeding behavior, aggression, and the response to stress.
Authors
JournalNeuron
PubMed ID18817733
Bone marrow sinusoidal endothelium controls terminal erythroid differentiation and reticulocyte maturation.
Authors
JournalNat Commun
PubMed ID34845225
An algorithm-based topographical biomaterials library to instruct cell fate.
Authors
JournalProc Natl Acad Sci U S A
PubMed ID21949368
An In Vivo CRISPR Screen Identifies Stepwise Genetic Dependencies of Metastatic Progression.
Authors
JournalCancer Res
PubMed ID34916221
A CDC42EP4/septin-based perisynaptic glial scaffold facilitates glutamate clearance.
Authors
JournalNat Commun
PubMed ID26657011
Acquisition of innate odor preference depends on spontaneous and experiential activities during critical period.
Authors
JournalElife
PubMed ID33769278
Circulating Tumor Cell Clustering Shapes DNA Methylation to Enable Metastasis Seeding.
Authors
JournalCell
PubMed ID30633912
PPARa-mediated peroxisome induction compensates PPAR?-deficiency in bronchiolar club cells.
AuthorsKarnati S, Oruqaj G, Janga H, Tumpara S, Colasante C, Van Veldhoven PP, Braverman N, Pilatz A, Mariani TJ, Baumgart-Vogt E
JournalPLoS One
PubMed ID30212482
Despite the important functions of PPAR? in various cell types of the lung, PPAR?-deficiency in club cells induces only mild emphysema. Peroxisomes are distributed in a similar way as PPAR? in the lung and are mainly enriched in club and AECII cells. To date, the effects of PPAR?-deficiency on the ... More
Bsx controls pineal complex development.
AuthorsSchredelseker T, Driever W
JournalDevelopment
PubMed ID29945867
Neuroendocrine cells in the pineal gland release melatonin during the night and, in teleosts, are directly photoreceptive. During development of the pineal complex, a small number of cells migrate leftward away from the pineal anlage to form the parapineal cell cluster, a process that is crucial for asymmetrical development of ... More
Identification of matrix physicochemical properties required for renal epithelial cell tubulogenesis by using synthetic hydrogels.
AuthorsCruz-Acuña R, Mulero-Russe A, Clark AY, Zent R, García AJ
JournalJ Cell Sci
PubMed ID31558679
Synthetic hydrogels with controlled physicochemical matrix properties serve as powerful
Dopamine inhibits human CD8+ Treg function through D
AuthorsNasi G, Ahmed T, Rasini E, Fenoglio D, Marino F, Filaci G, Cosentino M
JournalJ Neuroimmunol
PubMed ID30954278
CD8+ T regulatory/suppressor cells (Treg) affect peripheral tolerance and may be involved in autoimmune diseases as well as in cancer. In view of our previous data showing the ability of DA to affect adaptive immune responses, we investigated the dopaminergic phenotype of human CD8+ Treg as well as the ability ... More
Super-resolution imaging reveals the evolution of higher-order chromatin folding in early carcinogenesis.
AuthorsXu J, Ma H, Ma H, Jiang W, Mela CA, Duan M, Zhao S, Gao C, Hahm ER, Lardo SM, Troy K, Sun M, Pai R, Stolz DB, Zhang L, Singh S, Brand RE, Hartman DJ, Hu J, Hainer SJ, Liu Y
JournalNat Commun
PubMed ID32313005
Genomic DNA is folded into a higher-order structure that regulates transcription and maintains genomic stability. Although progress has been made on understanding biochemical characteristics of epigenetic modifications in cancer, the in-situ higher-order folding of chromatin structure during malignant transformation remains largely unknown. Here, using optimized stochastic optical reconstruction microscopy (STORM) for ... More
Targeting chemoresistant colorectal cancer via systemic administration of a BMP7 variant.
AuthorsVeschi V, Mangiapane LR, Nicotra A, Di Franco S, Scavo E, Apuzzo T, Sardina DS, Fiori M, Benfante A, Colorito ML, Cocorullo G, Giuliante F, Cipolla C, Pistone G, Bongiorno MR, Rizzo A, Tate CM, Wu X, Rowlinson S, Stancato LF, Todaro M, De Maria R, Stassi G
JournalOncogene
PubMed ID31591478
Despite intense research and clinical efforts, patients affected by advanced colorectal cancer (CRC) have still a poor prognosis. The discovery of colorectal (CR) cancer stem cell (CSC) as the cell compartment responsible for tumor initiation and propagation may provide new opportunities for the development of new therapeutic strategies. Given the ... More