A comparison between different human hepatocyte models reveals profound differences in net glucose production, lipid composition and metabolism in vitro.
AuthorsBonanini F,Singh M,Yang H,Kurek D,Harms AC,Mardinoglu A,Hankemeier T
JournalExperimental cell research
PubMed ID38499143
Effects of an indole derivative on cell proliferation, transfection, and alternative splicing in production of lentiviral vectors by transient co-transfection.
AuthorsMier NC,Roper DK
JournalPloS one
PubMed ID38833479
Lentiviral vectors derived from human immunodeficiency virus type I are widely used to deliver functional gene copies to mammalian cells for research and gene therapies. Post-transcriptional splicing of lentiviral vector transgene in transduced host and transfected producer cells presents barriers to widespread application of lentiviral vector-based therapies. The present study ... More
Point mutations identified in Lec8 Chinese hamster ovary glycosylation mutants that inactivate both the UDP-galactose and CMP-sialic acid transporters.
AuthorsOelmann S, Stanley P, Gerardy-Schahn R,
JournalJ Biol Chem
PubMed ID11319223
'Nucleotide-sugar transporters (NSTs) are critical components of glycosylation pathways in eukaryotes. The identification of structural elements that are involved in NST functions provides an important task. Chinese hamster ovary glycosylation mutants defective in nucleotide-sugar transport provide access to inactive transporters that can define such structure/function relationships. In this study, we ... More
The generation of rhythmic activity in dissociated cultures of rat spinal cord.
AuthorsStreit J; Tscherter A; Heuschkel MO; Renaud P
JournalEur J Neurosci
PubMed ID11553272
'Locomotion in vertebrates is controlled by central pattern generators in the spinal cord. The roles of specific network architecture and neuronal properties in rhythm generation by such spinal networks are not fully understood. We were able to induce similar patterns of rhythmic activity in dissociated cultures as in slice ... More
Production, concentration and titration of pseudotyped HIV-1-based lentiviral vectors.
AuthorsKutner RH, Zhang XY, Reiser J,
JournalNat Protoc
PubMed ID19300443
'Over the past decade, lentiviral vectors have emerged as powerful tools for transgene delivery. The use of lentiviral vectors has become commonplace and applications in the fields of neuroscience, hematology, developmental biology, stem cell biology and transgenesis are rapidly emerging. Also, lentiviral vectors are at present being explored in the ... More
Oligomerization of the human serotonin transporter and of the rat GABA transporter 1 visualized by fluorescence resonance energy transfer microscopy in living cells.
AuthorsSchmid JA, Scholze P, Kudlacek O, Freissmuth M, Singer EA, Sitte HH,
JournalJ Biol Chem
PubMed ID11071889
'Recent biochemical studies indicate that the serotonin transporter can form oligomers. We investigated whether the human serotonin transporter (hSERT) can be visualized as an oligomer in the plasma membrane of intact cells. For this purpose, we generated fusion proteins of hSERT and spectral variants of the green fluorescent protein (cyan ... More
Survival motor neuron protein modulates neuron-specific apoptosis.
AuthorsKerr DA, Nery JP, Traystman RJ, Chau BN, Hardwick JM,
JournalProc Natl Acad Sci U S A
PubMed ID11078511
'Spinal muscular atrophy (SMA) is attributed to mutations in the SMN1 gene, leading to loss of spinal cord motor neurons. The neurotropic Sindbis virus vector system was used to investigate a role for the survival motor neuron (SMN) protein in regulating neuronal apoptosis. Here we show that SMN protects primary ... More
D2 receptors inhibit the secretory process downstream from calcium influx in dopaminergic neurons: implication of K+ channels.
AuthorsCongar P; Bergevin A; Trudeau LE
JournalJ Neurophysiol
PubMed ID11826068
'Dopaminergic (DAergic) neurons possess D2-like somatodendritic and terminal autoreceptors that modulate cellular excitability and dopamine (DA) release. The cellular and molecular processes underlying the rapid presynaptic inhibition of DA release by D2 receptors remain unclear. Action-potential evoked EPSCs were reversibly inhibited by quinpirole, a selective D2 receptor agonist. This ... More
Gene correction of the apolipoprotein (Apo) E2 phenotype to wild-type ApoE3 by in situ chimeraplasty.
AuthorsTagalakis AD, Graham IR, Riddell DR, Dickson JG, Owen JS,
JournalJ Biol Chem
PubMed ID11278248
'Apolipoprotein (apo) E is a polymorphic plasma protein, synthesized mainly by liver. Here, we evaluate whether synthetic DNA-RNA oligonucleotides (chimeraplasts) can convert a dysfunctional isoform, apoE2 (C --> T, R158C), which causes Type III hyperlipidemia and premature atherosclerosis, into apoE3. First, we treated recombinant Chinese hamster ovary cells stably secreting ... More
A novel site on the Galpha -protein that recognizes heptahelical receptors.
'Specific domains of the G-protein alpha subunit have been shown to control coupling to heptahelical receptors. The extreme N and C termini and a region between alpha4 and alpha5 helices of the G-protein alpha subunit are known to determine selective interaction with the receptors. The metabotropic glutamate receptor 2 activated ... More
Co-regulation of survival of motor neuron (SMN) protein and its interactor SIP1 during development and in spinal muscular atrophy.
AuthorsJablonka S, Bandilla M, Wiese S, Bühler D, Wirth B, Sendtner M, Fischer U,
JournalHum Mol Genet
PubMed ID11181573
'Spinal muscular atrophy (SMA) is a neuromuscular disease characterized by the degeneration of motor neurons in the spinal cord. The disease is caused by mutations of the survival of motor neuron 1 gene (SMN1), resulting in a reduced production of functional SMN protein. A major question unanswered thus far is ... More
The Role of Constant Region Carbohydrate in the Assembly and Secretion of Human IgD and IgA1.
Authors Gala Francoise A; Morrison Sherie L;
JournalJ Biol Chem
PubMed ID12023968
'Immunoglobulins are glycoproteins, containing N- linked carbohydrates in the heavy chain constant regions of all isotypes and O-linked carbohydrates in the hinge regions of human IgA1 and IgD. A previous study showed that IgD synthesized in the presence of tunicamycin and lacking the three N-linked glycans on the heavy chain ... More
Activation of the Drosophila NF-kappaB factor Relish by rapid endoproteolytic cleavage.
AuthorsStöven S, Ando I, Kadalayil L, Engström Y, Hultmark D,
JournalEMBO Rep
PubMed ID11269501
The Rel/NF-kappaB transcription factor Relish plays a key role in the humoral immune response in Drosophila. We now find that activation of this innate immune response is preceded by rapid proteolytic cleavage of Relish into two parts. An N-terminal fragment, containing the DNA-binding Rel homology domain, translocates to the nucleus ... More
The sortilin cytoplasmic tail conveys Golgi-endosome transport and binds the VHS domain of the GGA2 sorting protein.
AuthorsNielsen MS, Madsen P, Christensen EI, Nykjaer A, Gliemann J, Kasper D, Pohlmann R, Petersen CM,
JournalEMBO J
PubMed ID11331584
Sortilin belongs to a growing family of multiligand type-1 receptors with homology to the yeast receptor Vps10p. Based on structural features and sortilin's intracellular predominance, we have proposed it to be a sorting receptor for ligands in the synthetic pathway as well as on the cell membrane. To test this ... More
A novel cytosolic dual specificity phosphatase, interacting with glucokinase, increases glucose phosphorylation rate.
AuthorsMuñoz-Alonso MJ, Guillemain G, Kassis N, Girard J, Burnol AF, Leturque A,
JournalJ Biol Chem
PubMed ID10913113
A novel protein was cloned from a rat liver cDNA library by interaction with the liver glucokinase. This protein contained 339 residues and possessed a canonical consensus sequence for a dual specificity phosphatase. The recombinant protein was able to dephosphorylate phosphotyrosyl and phosphoseryl/threonyl substrates. We called this protein the glucokinase-associated ... More
Molecular analysis of gene expression in the developing pontocerebellar projection system.
AuthorsDÃaz E, Ge Y, Yang YH, Loh KC, Serafini TA, Okazaki Y, Hayashizaki Y, Speed TP, Ngai J, Scheiffele P,
JournalNeuron
PubMed ID12408845
As an approach toward understanding the molecular mechanisms of neuronal differentiation, we utilized DNA microarrays to elucidate global patterns of gene expression during pontocerebellar development. Through this analysis, we identified groups of genes specific to neuronal precursor cells, associated with axon outgrowth, and regulated in response to contact with synaptic ... More
IDH mutation impairs histone demethylation and results in a block to cell differentiation.
Recurrent mutations in isocitrate dehydrogenase 1 (IDH1) and IDH2 have been identified in gliomas, acute myeloid leukaemias (AML) and chondrosarcomas, and share a novel enzymatic property of producing 2-hydroxyglutarate (2HG) from a-ketoglutarate. Here we report that 2HG-producing IDH mutants can prevent the histone demethylation that is required for lineage-specific progenitor ... More
Insulin and insulin-like growth factor I receptors utilize different G protein signaling components.
We examined the role of heterotrimeric G protein signaling components in insulin and insulin-like growth factor I (IGF-I) action. In HIRcB cells and in 3T3L1 adipocytes, treatment with the Galpha(i) inhibitor (pertussis toxin) or microinjection of the Gbetagamma inhibitor (glutathione S-transferase-betaARK) inhibited IGF-I and lysophosphatidic acid-stimulated mitogenesis but had no ... More
Characterization of pluripotent stem cells.
AuthorsMartí M, Mulero L, Pardo C, Morera C, Carrió M, Laricchia-Robbio L, Esteban CR, Izpisua Belmonte JC,
JournalNat Protoc
PubMed ID23306458
Characterization of pluripotent stem cells is required for the registration of stem cell lines and allows for an impartial and objective comparison of the results obtained when generating multiple lines. It is therefore crucial to establish specific, fast and reliable protocols to detect the hallmarks of pluripotency. Such protocols should ... More
Polycystin-1, the product of the polycystic kidney disease 1 gene, co-localizes with desmosomes in MDCK cells.
AuthorsScheffers MS, van der Bent P, Prins F, Spruit L, Breuning MH, Litvinov SV, de Heer E, Peters DJ,
JournalHum Mol Genet
PubMed ID11063733
Polycystin-1 is a novel protein predicted to be a large membrane-spanning glycoprotein with an extracellular N-terminus and an intracellular C-terminus, harboring several structural motifs. To study the subcellular localization, antibodies raised against various domains of polycystin-1 and against specific adhesion complex proteins were used for two-color immunofluorescence staining. In Madine ... More
Efficient intracellular assembly of papillomaviral vectors.
Although the papillomavirus structural proteins, L1 and L2, can spontaneously coassemble to form virus-like particles, currently available methods for production of L1/L2 particles capable of transducing reporter plasmids into mammalian cells are technically demanding and relatively low-yield. In this report, we describe a simple 293 cell transfection method for efficient ... More
Calcium and calpain as key mediators of apoptosis-like death induced by vitamin D compounds in breast cancer cells.
AuthorsMathiasen IS, Sergeev IN, Bastholm L, Elling F, Norman AW, Jaattela M.
JournalJ Biol Chem
PubMed ID12072431
The active form of vitamin D(3) (1,25(OH)(2)D(3)) induces an increase in the intracellular free calcium ([Ca(2+)](i)) and caspase-independent cell death in human breast cancer cells. Here we show that the treatment of MCF-7 breast cancer cells with 1,25(OH)(2)D(3) or its chemotherapeutic analog, EB 1089, releases Ca(2+) from the endoplasmic reticulum. ... More
Alzheimer's disease-related overexpression of the cation-dependent mannose 6-phosphate receptor increases Abeta secretion: role for altered lysosomal hydrolase distribution in beta-amyloidogenesis.
Authors Mathews Paul M; Guerra Carolyn B; Jiang Ying; Grbovic Olivera M; Kao Benjamin H; Schmidt Stephen D; Dinakar Ravi; Mercken Marc; Hille-Rehfeld Annette; Rohrer Jack; Mehta Pankaj; Cataldo Anne M; Nixon Ralph A;
JournalJ Biol Chem
PubMed ID11551970
Prominent endosomal and lysosomal changes are an invariant feature of neurons in sporadic Alzheimer's disease (AD). These changes include increased levels of lysosomal hydrolases in early endosomes and increased expression of the cation-dependent mannose 6-phosphate receptor (CD-MPR), which is partially localized to early endosomes. To determine whether AD-associated redistribution of ... More