Endogenous endothelial cell nitric-oxide synthase modulates apoptosis in cultured breast cancer cells and is transcriptionally regulated by p53.
AuthorsMortensen K,Skouv J,Hougaard DM,Larsson LI
JournalThe Journal of biological chemistry
PubMed ID10608825
5-methyltetrahydrofolate, the active form of folic acid, restores endothelial function in familial hypercholesterolemia.
AuthorsVerhaar MC, Wever RM, Kastelein JJ, van Dam T, Koomans HA, Rabelink TJ
JournalCirculation
PubMed ID9462523
BACKGROUND: Impaired nitric oxide (NO) activity is an early event in the pathogenesis of cardiovascular disease, resulting from either reduced NO formation or increased NO degradation. Administration of tetrahydrobiopterin (BH4), an essential cofactor for NO production, could restore NO activity in familial hypercholesterolemia (FH). Because folates have been suggested to ... More
Sarco/endoplasmic reticulum calcium ATPase-2 expression is regulated by ATF6 during the endoplasmic reticulum stress response: intracellular signaling of calcium stress in a cardiac myocyte model system.
AuthorsThuerauf DJ, Hoover H, Meller J, Hernandez J, Su L, Andrews C, Dillmann WH, McDonough PM, Glembotski CC
JournalJ Biol Chem
PubMed ID11595740
The recently described transcription factor, ATF6, mediates the expression of proteins that compensate for potentially stressful changes in the endoplasmic reticulum (ER), such as reduced ER calcium. In cardiac myocytes the maintenance of optimal calcium levels in the sarcoplasmic reticulum (SR), a specialized form of the ER, is required for ... More
A novel zinc finger protein TReP-132 interacts with CBP/p300 to regulate human CYP11A1 gene expression.
AuthorsGizard F, Lavallée B, DeWitte F, Hum DW
JournalJ Biol Chem
PubMed ID11349124
'The human CYP11A1 gene is expressed specifically in steroidogenic tissues and encodes cytochrome P450scc, which catalyzes the first step in steroid synthesis. A region of the 5'-flanking DNA of the gene from nucleotides -155 to -131 (-155/-131) is shown to activate transcription in steroidogenic human placental JEG-3 (1) and adrenal ... More
Tec kinase signaling in T cells is regulated by phosphatidylinositol 3-kinase and the Tec pleckstrin homology domain.
AuthorsYang WC, Ching KA, Tsoukas CD, Berg LJ
JournalJ Immunol
PubMed ID11123316
'Tec, the prototypical member of the Tec family of tyrosine kinases, is abundantly expressed in T cells and other hemopoietic cell types. Although the functions of Itk and Txk have recently been investigated, little is known about the role of Tec in T cells. Using antisense oligonucleotide treatment to deplete ... More
A role for the p38 mitogen-activated protein kinase pathway in myocardial cell growth, sarcomeric organization, and cardiac-specific gene expression.
AuthorsZechner D, Thuerauf DJ, Hanford DS, McDonough PM, Glembotski CC
JournalJ Cell Biol
PubMed ID9314533
'Three hallmark features of the cardiac hypertrophic growth program are increases in cell size, sarcomeric organization, and the induction of certain cardiac-specific genes. All three features of hypertrophy are induced in cultured myocardial cells by alpha1- adrenergic receptor agonists, such as phenylephrine (PE) and other growth factors that activate mitogen- ... More
Involvement of nuclear factor of activated T cells activation in UV response. Evidence from cell culture and transgenic mice.
AuthorsHuang C, Mattjus P, Ma WY, Rincon M, Chen NY, Brown RE, Dong Z
JournalJ Biol Chem
PubMed ID10734048
'Mammalian cells respond to UV radiation by signaling cascades leading to activation of transcription factors, such as activated protein 1, NFkappaB, and p53, a process known as the "UV response." Nuclear factor of activated T cells (NFAT) was first identified as an inducible nuclear factor in immune response and subsequently ... More
Activation of nuclear factor-kappaB is necessary for myotrophin-induced cardiac hypertrophy.
AuthorsGupta S, Purcell NH, Lin A, Sen S
JournalJ Cell Biol
PubMed ID12486112
'The transcription factor nuclear factor-kappaB (NF-kappaB) regulates expression of a variety of genes involved in immune responses, inflammation, proliferation, and programmed cell death (apoptosis). Here, we show that in rat neonatal ventricular cardiomyocytes, activation of NF-kappaB is involved in the hypertrophic response induced by myotrophin, a hypertrophic activator identified from ... More
N-cadherin-dependent cell-cell contact regulates Rho GTPases and beta-catenin localization in mouse C2C12 myoblasts.
AuthorsCharrasse S, Meriane M, Comunale F, Blangy A, Gauthier-Rouvière C
JournalJ Cell Biol
PubMed ID12213839
'N-cadherin, a member of the Ca(2+)-dependent cell-cell adhesion molecule family, plays an essential role in skeletal muscle cell differentiation. We show that inhibition of N-cadherin-dependent adhesion impairs the upregulation of the two cyclin-dependent kinase inhibitors p21 and p27, the expression of the muscle-specific genes myogenin and troponin T, and C2C12 ... More
1,25-Dihydroxyvitamin D down-regulates cell membrane growth- and nuclear growth-promoting signals by the epidermal growth factor receptor.
AuthorsCordero JB, Cozzolino M, Lu Y, Vidal M, Slatopolsky E, Stahl PD, Barbieri MA, Dusso A
JournalJ Biol Chem
PubMed ID12181310
'1,25(OH)(2)D(3) antiproliferative properties are widely known. However, the molecular bases of these properties are only partially elucidated. Since 1,25(OH)(2)D(3) effectively arrests growth in many tumors and hyperplastic tissues whose growth is driven by co-expression of EGFR and its ligand TGF-alpha, it was hypothesized that 1,25(OH)(2)D(3) could affect the TGF-alpha/EGFR-autocrine growth ... More
Regulation and reversibility of vacuolar H(+)-ATPase.
AuthorsHirata T, Nakamura N, Omote H, Wada Y, Futai M
JournalJ Biol Chem
PubMed ID10617629
'Arabidopsis thaliana vacuolar H(+)-translocating pyrophosphatase (V-PPase) was expressed functionally in yeast vacuoles with endogenous vacuolar H(+)-ATPase (V-ATPase), and the regulation and reversibility of V-ATPase were studied using these vacuoles. Analysis of electrochemical proton gradient (DeltamuH) formation with ATP and pyrophosphate indicated that the proton transport by V-ATPase or V-PPase is ... More
CARD11 and CARD14 are novel caspase recruitment domain (CARD)/membrane-associated guanylate kinase (MAGUK) family members that interact with BCL10 and activate NF-kappa B.
AuthorsBertin J, Wang L, Guo Y, Jacobson MD, Poyet JL, Srinivasula SM, Merriam S, DiStefano PS, Alnemri ES
JournalJ Biol Chem
PubMed ID11278692
'The caspase recruitment domain (CARD) is a protein-binding module that mediates the assembly of CARD-containing proteins into apoptosis and NF-kappaB signaling complexes. We report here that CARD protein 11 (CARD11) and CARD protein 14 (CARD14) are novel CARD-containing proteins that belong to the membrane-associated guanylate kinase (MAGUK) family, a class ... More
The transcriptional coactivator CREB-binding protein cooperates with STAT1 and NF-kappa B for synergistic transcriptional activation of the CXC ligand 9/monokine induced by interferon-gamma gene.
AuthorsHiroi M, Ohmori Y
JournalJ Biol Chem
PubMed ID12403783
'Signal transducers and activators of transcription 1 (STAT1) and NF-kappaB cooperatively regulate the expression of many inflammatory genes. In the present study, we demonstrate that the transcriptional coactivator CREB-binding protein (CBP) mediated the STAT1/NF-kappaB synergy for transcription of the gene for CXC ligand 9 (CXCL9), an interferon-gamma (IFN-gamma)-inducible chemokine. Reporter ... More
Gonadotropin depression of adenosine triphosphate levels and interaction with adenosine in rat granulosa cells.
AuthorsBillig H, Rosberg S
JournalEndocrinology
PubMed ID3002759
'Cellular ATP levels were measured with the luceferin-luciferase enzyme method in incubated preovulatory granulosa cells in vitro from PMSG-treated immature rats. The ATP levels were depressed by both FSH and LH, FSH being the more effective. Adenosine enhanced the ATP levels about 3-fold, but the depressive effects of gonadotropins could ... More
CCD imaging of luciferase gene expression in single mammalian cells.
AuthorsHooper CE, Ansorge RE, Browne HM, Tomkins P
JournalJ Biolumin Chemilumin
PubMed ID2336972
'Quantitative and sensitive imaging of chemiluminescence, bioluminescence and fluorescence emissions is emerging as an increasingly important technique for a range of biomedical applications (Hooper et al., 1990). A brief review of low-light-level imaging is presented, with particular reference to charge-coupled devices (CCD). Detectors for sensitive imaging are described and compared, ... More
Cyclic AMP- and cyclic GMP-dependent protein kinases differ in their regulation of cyclic AMP response element-dependent gene transcription.
AuthorsCollins SP, Uhler MD
JournalJ Biol Chem
PubMed ID10085070
'The ability of cGMP-dependent protein kinases (cGKs) to activate cAMP response element (CRE)-dependent gene transcription was compared with that of cAMP-dependent protein kinases (cAKs). Although both the type Ibeta cGMP-dependent protein kinase (cGKIbeta) and the type II cAMP-dependent protein kinase (cAKII) phosphorylated the cytoplasmic substrate VASP (vasodilator- and A kinase-stimulated ... More
Diaminobiotin and desthiobiotin have biotin-like activities in Jurkat cells.
AuthorsRodriguez-Melendez R, Lewis B, McMahon RJ, Zempleni J
JournalJ Nutr
PubMed ID12730407
'In mammals, biotin serves as a coenzyme for carboxylases such as propionyl-CoA carboxylase. The expression of genes encoding interleukin-2 (IL-2) and IL-2 receptor (IL-2R)gamma also depends on biotin. Biotin metabolites are structurally similar to biotin, and their concentrations in tissues are quantitatively important. Here, the hypothesis was tested that biotin ... More
Comparison of MTT and ATP-based assays for the measurement of viable cell number.
AuthorsPetty RD, Sutherland LA, Hunter EM, Cree IA
JournalJ Biolumin Chemilumin
PubMed ID7762413
'Cell viability assays are widely used to assess the effect chemotherapeutic drugs and other agents on cell lines and have shown promise for the prediction of tumour chemosensitivity. In this study we have compared two viability assays using Daudi and CCRF-CEM cell lines over a range of 1500-100,000 cells/well of ... More
Long-term monitoring of circadian rhythms in c-fos gene expression from suprachiasmatic nucleus cultures.
AuthorsGeusz ME, Fletcher C, Block GD, Straume M, Copeland NG, Jenkins NA, Kay SA, Day RN
JournalCurr Biol
PubMed ID9368758
'BACKGROUND: The AP-1 family of transcription factors has been implicated in the control of the expression of many genes in response to environmental signals. Previous studies have provided temporal profiles for c-fos expression by taking measurements from many animals at several points in time, but these studies provide limited information ... More
G alpha 16 couples chemoattractant receptors to NF-kappa B activation.
AuthorsYang M, Sang H, Rahman A, Wu D, Malik AB, Ye RD
JournalJ Immunol
PubMed ID11359849
'The guanine nucleotide-binding regulatory protein alpha-subunit, Galpha(16), is primarily expressed in hemopoietic cells, and interacts with a large number of seven-membrane span receptors including chemoattractant receptors. We investigated the biological functions resulting from Galpha(16) coupling of chemoattractant receptors in a transfected cell model system. HeLa cells expressing a kappaB-driven luciferase ... More
Functional interactions of lanthanum and phospholipase D with the abscisic acid signaling effectors VP1 and ABI1-1 in rice protoplasts.
AuthorsGampala SS, Hagenbeek D, Rock CD
JournalJ Biol Chem
PubMed ID11139577
'cis,trans-Abscisic acid (ABA) plays an important role in plant growth and development, regulation of seed maturation, germination, and adaptation to environmental stresses. Knowledge of ABA mechanisms of action and the interactions of components required for ABA signal transduction is far from complete. Using transient gene expression in rice protoplasts, we ... More
Cytoplasmic factors that affect the intensity and stability of bioluminescence from firefly luciferase in living mammalian cells.
AuthorsGandelman O, Allue I, Bowers K, Cobbold P
JournalJ Biolumin Chemilumin
PubMed ID7879652
'In order to improve calibration of firefly luciferase signals obtained by injecting the enzyme into single, isolated heart and liver cells we have investigated why the luminescence from cells is greatly depressed compared with in vitro (in mammalian ionic milieu) and why the decay of the intracellular signal is remarkably ... More
Cloning and expression of cDNA for the luciferase from the marine ostracod Vargula hilgendorfii.
AuthorsThompson EM, Nagata S, Tsuji FI
JournalProc Natl Acad Sci U S A
PubMed ID2771943
'The marine ostracod Vargula hilgendorfii ejects luciferin and luciferase into seawater to produce a bright luminous cloud. The light is due to the oxidation of luciferin, an imidazopyrazine compound, by molecular oxygen, catalyzed by luciferase. The mechanism of the reaction has been studied extensively and the 60 kcal/mol required for ... More
Surface expression and endocytosis of the human cytomegalovirus-encoded chemokine receptor US28 is regulated by agonist-independent phosphorylation.
AuthorsMokros T, Rehm A, Droese J, Oppermann M, Lipp M, Höpken UE
JournalJ Biol Chem
PubMed ID12244063
'Human cytomegalovirus encodes the G protein-coupled chemokine receptor homologue US28 that binds several CC chemokines and sequesters extracellular chemokines from the environment of infected cells. Mechanistically, it has been shown that US28 undergoes rapid constitutive receptor endocytosis and recycling. Monoclonal antibodies were raised that allowed the characterization of a ligand-independent ... More
Parathyroid hormone potentiates nucleotide-induced [Ca2+]i release in rat osteoblasts independently of Gq activation or cyclic monophosphate accumulation. A mechanism for localizing systemic responses in bone.
AuthorsBuckley KA, Wagstaff SC, McKay G, Gaw A, Hipskind RA, Bilbe G, Gallagher JA, Bowler WB
JournalJ Biol Chem
PubMed ID11124938
'The regulation of tissue turnover requires the coordinated activity of both local and systemic factors. Nucleotides exist transiently in the extracellular environment, where they serve as ligands to P2 receptors. Here we report that the localized release of these nucleotides can sensitize osteoblasts to the activity of systemic factors. We ... More
Specific subdomains of Vav differentially affect T cell and NK cell activation.
'The Vav protooncogene is a multidomain protein involved in the regulation of IL-2 gene transcription in T cells and the development of cell-mediated killing by cytotoxic lymphocytes. We have investigated the differential roles that specific protein subdomains within the Vav protooncogene have in the development of these two distinct cellular ... More
Modulation of cell signaling pathways can enhance or impair glucocorticoid-induced gene expression without altering the state of receptor phosphorylation.
AuthorsMoyer ML, Borror KC, Bona BJ, DeFranco DB, Nordeen SK
JournalJ Biol Chem
PubMed ID7693681
'We have stably introduced expression vectors for the glucocorticoid receptor and a sensitive, hormone-responsive reporter (mouse mammary tumor virus-luciferase) into a human breast carcinoma-derived cell line. Employing this cell line, we have conducted a detailed examination of the induction of glucocorticoid-regulated genes and the phosphorylation of glucocorticoid receptor following pharmacologic ... More
Variable patterns of expression of luciferase in transgenic tobacco leaves.
AuthorsBarnes WM
JournalProc Natl Acad Sci U S A
PubMed ID2251262
'A carboxyl-terminally modified firefly luciferase, encoded as a gene fusion to the neomycin phosphotransferase gene (which confers kanamycin resistance), was found to be enzymatically active for both enzymes when expressed in bacteria and in transgenic plants. A military-type starlight vision system was used to conveniently analyze the pattern of gene ... More
A novel circadian phenotype based on firefly luciferase expression in transgenic plants.
AuthorsMillar AJ, Short SR, Chua NH, Kay SA
JournalPlant Cell
PubMed ID1392609
'A 320-bp fragment of the Arabidopsis cab2 promoter is sufficient to mediate transcriptional regulation by both phytochrome and the circadian clock. We fused this promoter fragment to the firefly luciferase (Luc) gene to create a real-time reporter for regulated gene expression in intact plants. Cab2::Luc transcript accumulated in the expected ... More
Membrane-permeable luciferin esters for assay of firefly luciferase in live intact cells.
AuthorsCraig FF, Simmonds AC, Watmore D, McCapra F, White MR
JournalBiochem J
PubMed ID2064603
'Five esters of luciferin were synthesized and compared with native luciferin as substrates for firefly luciferase expressed in live intact mammalian cells. The esters themselves were not substrates for purified luciferase, but four were substrates for a purified esterase and all appeared to be hydrolysed to luciferin within mammalian cells. ... More
Secretagogues modulate the calcium concentration in the endoplasmic reticulum of insulin-secreting cells. Studies in aequorin-expressing intact and permeabilized ins-1 cells.
AuthorsMaechler P, Kennedy ED, Sebö E, Valeva A, Pozzan T, Wollheim CB
JournalJ Biol Chem
PubMed ID10212237
'The precise regulation of the Ca2+ concentration in the endoplasmic reticulum ([Ca2+]er) is important for protein processing and signal transduction. In the pancreatic beta-cell, dysregulation of [Ca2+]er may cause impaired insulin secretion. The Ca2+-sensitive photoprotein aequorin mutated to lower its Ca2+ affinity was stably expressed in the endoplasmic reticulum (ER) ... More
Fast charge translocations associated with partial reactions of the Na,K-pump: I. Current and voltage transients after photochemical release of ATP.
AuthorsBorlinghaus R, Apell HJ, Läuger P
JournalJ Membr Biol
PubMed ID3041002
'Nonstationary electric currents are described which are generated by the Na,K-pump. Flat membrane sheets 0.2-1 micron in diameter containing a high density of oriented Na,K-ATPase molecules are bound to a planar lipid bilayer acting as a capacitive electrode. In the aqueous phase adjacent to the bound membrane sheets, ATP is ... More
Transcriptional regulation of P450scc gene expression in the embryonic rodent nervous system.
AuthorsHammer F, Compagnone NA, Vigne JL, Bair SR, Mellon SH
JournalEndocrinology
PubMed ID14576192
'Steroid hormones are synthesized in adrenals, gonads, placenta, and the central and peripheral nervous systems (neurosteroids). Neurosteroidogenesis, like conventional steroidogenesis, begins with the conversion of cholesterol to pregnenolone, catalyzed by mitochondrial P450 side-chain cleavage enzyme (P450scc). Transcription of the P450scc gene in the adrenals and gonads requires steroidogenic factor-1, which ... More
Discovery and validation of a new family of antioxidants: the aminopyrazine derivatives.
'Coelenteramine (2-amino-1,4-pyrazine derivative), one of the metabolites of the oxidative degradation of coelenterazine (imidazolopyrazinone derivative), is endowed with excellent antioxidative properties towards ROS/RNS, like its mother-compound. This crucial discovery, made during the study of natural bioluminescent compounds (luciferins), has stimulated the development of synthetic aminopyrazine derivatives as new leads in ... More
A peroxidase-independent method for the quantitation of extracellular hydrogen peroxide generated by activated phagocytes in vitro.
AuthorsAnderson R
JournalJ Immunol Methods
PubMed ID1328397
'A peroxidase-independent method for the determination of phagocyte-derived hydrogen peroxide in vitro is described. The method is based on the catalase-inhibitable oxidation of added cysteine. Human blood neutrophils (1 x 10(6)/ml) were coincubated with the myeloperoxidase (MPO) inhibitor, sodium azide (10 micrograms/ml), for 30 min at 37 degrees C followed ... More
Catalytic activities of mitochondrial ATP synthase in patients with mitochondrial DNA T8993G mutation in the ATPase 6 gene encoding subunit a.
AuthorsBaracca A, Barogi S, Carelli V, Lenaz G, Solaini G
JournalJ Biol Chem
PubMed ID10660580
'We investigated the biochemical phenotype of the mtDNA T8993G point mutation in the ATPase 6 gene, associated with neurogenic muscle weakness, ataxia, and retinitis pigmentosa (NARP), in three patients from two unrelated families. All three carried >80% mutant genome in platelets and were manifesting clinically various degrees of the NARP ... More
Targeting of cloned firefly luciferase to yeast mitochondria.
AuthorsAflalo C
JournalBiochemistry
PubMed ID2194561
'The firefly luciferase gene (luc) was fused to a 5'' fragment of the 70-kDa protein gene (70K) from yeast. The fragment codes for the N-terminal putative signal sequence which targets and anchors the 70-kDa protein to the cytoplasmic side of the outer membrane in mitochondria. Two versions of the fusion ... More
Establishment of a transient expression system for Dictyostelium discoideum.
AuthorsHoward PK, Ahern KG, Firtel RA
JournalNucleic Acids Res
PubMed ID3362676
'We have established a rapid and sensitive transient expression system for Dictyostelium discoideum. We constructed a gene fusion containing the promoter from the Dictyostelium Actin 15 gene fused to the firefly luciferase gene. The enzymatic activity of this gene fusion, expressed at very high levels in stable transformants, was measured ... More
Acetylcholine releases ATP from varicosities isolated from guinea pig myenteric plexus.
AuthorsWhite TD, Al-Humayyd M
JournalJ Neurochem
PubMed ID6403667
'The effects of cholinergic agonists and antagonists on the release of ATP from isolated myenteric varicosities were studied using a firefly luciferin-luciferase technique. In a previous study, acetylcholine and nicotine released ATP from isolated myenteric varicosities, whereas the muscarinic agonist bethanechol did not. In the present study, release of ATP ... More
Identification of nonmitochondrial creatine kinase enzymatic activity in isolated sea urchin mitotic apparatus.
AuthorsSilver RB, Saft MS, Taylor AR, Cole RD
JournalJ Biol Chem
PubMed ID6313691
'ATP-dependent calcium sequestration was previously localized in vesicles of mitotic apparatus isolated from sea urchins. We now demonstrate that the mitotic apparatus contains an ATP-regenerative system characterized as creatine kinase (EC 2.7.3.2). Mitotic apparatus isolated with vesicles intact converted ADP to ATP if phosphocreatine was present. Omission of ADP or ... More
The role of active site residue arginine 218 in firefly luciferase bioluminescence.
AuthorsBranchini BR, Magyar RA, Murtiashaw MH, Portier NC
JournalBiochemistry
PubMed ID11327861
'Firefly luciferase catalyzes the highly efficient emission of yellow-green light from substrate firefly luciferin by a sequence of reactions that require Mg-ATP and molecular oxygen. We had previously developed a working model of the luciferase active site based on the X-ray structure of the enzyme without bound substrates. In our ... More
Inhibition of tumor necrosis factor-alpha -induced nuclear translocation and activation of NF-kappa B by dehydroxymethylepoxyquinomicin.
AuthorsAriga A, Namekawa J, Matsumoto N, Inoue J, Umezawa K
JournalJ Biol Chem
PubMed ID11983688
'We previously designed and synthesized an NF-kappaB inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ), that showed anti-inflammatory activity in vivo. In the present study we looked into its mechanism of inhibition. DHMEQ inhibited tumor necrosis factor-alpha (TNF-alpha)- and 12-O-tetradecanoylphorbol-13-acetate-induced transcriptional activity of NF-kappaB in human T cell leukemia Jurkat cells. It also inhibited the ... More
Oxyluciferin, a luminescence product of firefly luciferase, is enzymatically regenerated into luciferin.
AuthorsGomi K, Kajiyama N
JournalJ Biol Chem
PubMed ID11457857
'The activity regenerating luciferin from the luminescent product oxyluciferin was found in the protein fraction of a lantern extract from Photinus pyralis. The protein, luciferin-regenerating enzyme (LRE), was purified to homogeneity by ammonium sulfate precipitation followed by successive column chromatography on Ultrogel AcA34, S-Sepharose FF, Q-Sepharose FF, TSKgel super Q ... More
CD28 utilizes Vav-1 to enhance TCR-proximal signaling and NF-AT activation.
AuthorsMichel F, Mangino G, Attal-Bonnefoy G, Tuosto L, Alcover A, Roumier A, Olive D, Acuto O
JournalJ Immunol
PubMed ID11034388
'The mechanism through which CD28 costimulation potentiates TCR-driven gene expression is still not clearly defined. Vav-1, an exchange factor for Rho GTPases thought to regulate, mainly through Rac-1, various signaling components leading to cytokine gene expression, is tyrosine phosphorylated upon CD28 engagement. Here, we provide evidence for a key role ... More
Firefly luciferase as a reporter enzyme for measuring gene expression in vegetative and symbiotic Rhizobium meliloti and other gram-negative bacteria.
AuthorsPalomares AJ, DeLuca MA, Helinski DR
JournalGene
PubMed ID2680767
'A DNA segment carrying a cDNA copy of the luciferase gene (luc) of the North American firefly Photinus pyralis, fused to the lambda PR promoter and expressed in Escherichia coli [de Wet et al., Proc. Natl. Acad. Sci. USA 82 (1985) 7870-7873], was inserted into a broad-host-range plasmid vector and ... More
Chemiluminescence microscopy as a tool in biomedical research.
AuthorsCréton R, Jaffe LF
JournalBiotechniques
PubMed ID11730016
'Chemiluminescence has become a standard tool in biomedical research. Chemiluminescent probes are used for immunoassays, nucleic acid identification, reporter gene assays, measuring enzyme activity, and the detection of ions and small molecules such as Ca2+, ATP, NO, O2- and H2O2. Along with the development of new chemiluminescent probes, significant progress ... More
Endothelin-1-specific activation of B-type natriuretic peptide gene via p38 mitogen-activated protein kinase and nuclear ETS factors.
AuthorsPikkarainen S, Tokola H, Kerkelä R, Majalahti-Palviainen T, Vuolteenaho O, Ruskoaho H
JournalJ Biol Chem
PubMed ID12446726
'Terminally differentiated cardiac myocytes adapt to mechanical and neurohumoral stress via morphological changes of individual cells accompanied by reactivation of fetal pattern of gene expression. Endothelin-1, a powerful paracrine mediator of myocyte growth, induces similar changes in cultured cardiac myocytes as those seen in hypertrophied heart in vivo. By using ... More
Evaluation of the proliferative response of lymphocytes by measurement of intracellular ATP.
AuthorsIshizaka A, Tono-oka T, Matsumoto S
JournalJ Immunol Methods
PubMed ID6431008
'The measurement of intracellular ATP levels by luciferin-luciferase-induced bioluminescence was used for the evaluation of proliferative responses of human mononuclear cells to lectins. A linear relationship was observed between the number of cells and the amount of ATP in the samples, and high reproducibility was obtained. The ATP content of ... More
The in vivo pattern of firefly luciferase expression in transgenic plants.
AuthorsSchneider M, Ow DW, Howell SH
JournalPlant Mol Biol
PubMed ID2102877
'Expression of the firefly luciferase gene in transgenic plants produces light emission patterns when the plants are supplied with luciferin. We explored whether in in vivo pattern of light emission truly reveals the pattern of luciferase gene expression or whether it reflects other parameters such as the availability of the ... More
Dictyostelium discoideum mutants with temperature-sensitive defects in endocytosis.
AuthorsBacon RA, Cohen CJ, Lewin DA, Mellman I
JournalJ Cell Biol
PubMed ID7929583
'We have isolated and characterized temperature-sensitive endocytosis mutants in Dictyostelium discoideum. Dictyostelium is an attractive model for genetic studies of endocytosis because of its high rates of endocytosis, its reliance on endocytosis for nutrient uptake, and tractable molecular genetics. Endocytosis-defective mutants were isolated by a fluorescence-activated cell sorting (FACS) as ... More
Cloning of firefly luciferase cDNA and the expression of active luciferase in Escherichia coli.
Authorsde Wet JR, Wood KV, Helinski DR, DeLuca M
JournalProc Natl Acad Sci U S A
PubMed ID3906652
'A cDNA library was constructed from firefly (Photinus pyralis) lantern poly(A)+ RNA, using the Escherichia coli expression vector lambda gt11. The library was screened with anti-P. pyralis luciferase (Photinus luciferin:oxygen 4-oxidoreductase, EC 1.13.12.7) antibody, and several cDNA clones expressing luciferase antigens were isolated. One clone, lambda Luc1, contained 1.5 kilobase ... More
Measurement of secretion of adenine nucleotides.
AuthorsHolmsen H, Dangelmaier CA
JournalMethods Enzymol
PubMed ID2716526
Mechanism of bioluminescence, chemiluminescence and enzyme function in the oxidation of firefly luciferin.
AuthorsMcElroy WD, Seliger HH, White EH
JournalPhotochem Photobiol
PubMed ID5824751
Utility of firefly luciferase as a reporter gene for promoter activity in transgenic mice.
AuthorsDiLella AG, Hope DA, Chen H, Trumbauer M, Schwartz RJ, Smith RG
JournalNucleic Acids Res
PubMed ID3375079
Introns increase gene expression in cultured maize cells.
AuthorsCallis J, Fromm M, Walbot V
JournalGenes Dev
PubMed ID2828168
'Using electroporation-mediated gene transfer, the gene encoding the Slow (S) migrating polypeptide of the maize (Zea mays L.) alcohol dehydrogenase-1 (Adh1) enzyme has been introduced stably and transiently into maize cells containing an endogenous Fast (F) ADH1 electromorph. In stable transformants an 11.5-kb fragment was sufficient to program normal S ... More
Synthesis of active firefly luciferase by in vitro translation of RNA obtained from adult lanterns.
AuthorsWood KV, de Wet JR, Dewji N, DeLuca M
JournalBiochem Biophys Res Commun
PubMed ID6208909
'Poly (A)+ RNA was isolated from the lanterns of adult fireflies, Photinus pyralis. The Poly (A)+ RNA was translated in a cell-free translation mixture from rabbit reticulocytes and the synthesis of enzymatically active firefly luciferase was demonstrated. The translation products were immunoprecipitated with anti-luciferase and then subjected to SDS gel ... More
Genetic analysis of the human tumor necrosis factor alpha/cachectin promoter region in a macrophage cell line.
'The 615-bp 5'' flanking region of the human TNF-alpha/cachectin gene was isolated and ligated to the luciferase reporter gene. In addition, a series of truncated promoter constructs was generated by exonuclease III digestion. The promoter activity of these constructs was studied in a transient transfection system using the TNF-alpha-producing U937 ... More
Novel monomeric luciferase enzymes as tools to study plant gene regulation in vivo.
AuthorsOlsson O, Nilsson O, Koncz C
JournalJ Biolumin Chemilumin
PubMed ID2186598
'Taking advantage of a specially constructed vector, luciferase LuxA and LuxB subunits were connected in frame to different amino acid linkers to reproduce a series of monomeric luciferase enzymes. A comparison of their activities in E. coli cells demonstrated that the length of the linkers positively affected activity. One luciferase ... More
Serum and insulin regulate cap function in 3T3-L1 cells.
AuthorsGallie DR, Traugh JA
JournalJ Biol Chem
PubMed ID8125928
'Serum starvation and readdition of serum, insulin, or mitogens result in changes in phosphorylation of several eukaryotic initiation factors that correlate with a decrease or stimulation in translation, respectively. We have introduced in vitro synthesized reporter mRNAs into exponentially growing and serum-deprived 3T3-L1 cells to examine whether serum deprivation affects ... More
An easily synthesized, photolyzable luciferase substrate for in vivo luciferase activity measurement.
AuthorsYang J, Thomason DB
JournalBiotechniques
PubMed ID8267979
'Many reporter gene assays require killing the cell by fixation or lysis. For assays in living cells, the substrate delivery is inefficient and cannot be supplied in situ in a bolus, which makes assays highly variable. We report a simple synthesis of a luciferin ester that is both photolyzable and ... More
Activation and cellular localization of the cyclosporine A-sensitive transcription factor NF-AT in skeletal muscle cells.
'The widely used immunosuppressant cyclosporine A (CSA) blocks nuclear translocation of the transcription factor, NF-AT (nuclear factor of activated T cells), preventing its activity. mRNA for several NF-AT isoforms has been shown to exist in cells outside of the immune system, suggesting a possible mechanism for side effects associated with ... More
Bioluminescence detection system of mutagen using firefly luciferase genes introduced in Escherichia coli lysogenic strain.
AuthorsLee SM, Suzuki M, Kumagai M, Ikeda H, Tamiya E, Karube I
JournalAnal Chem
PubMed ID1416034
'A rapid and convenient microbial sensing system for mutagens was developed based upon the induction of prophage from Escherichia coli lysogenic strain and bioluminescence. The system consisted of lysogenic E. coli encoding firefly luciferase genes and a photodetection system. Measurement of mutagen mitomycin C was achieved by measuring the luminescence ... More
Introduction to beetle luciferases and their applications.
AuthorsWood KV, Lam YA, McElroy WD
JournalJ Biolumin Chemilumin
PubMed ID2678917
'All beetle luciferases have evolved from a common ancestor: they all use ATP, O2, and a common luciferin as substrates. The most studied of these luciferases is that derived from the firefly Photinus pyralis, a beetle in the superfamily of Cantharoidea. The sensitivity with which the activity of this enzyme ... More
A potent new class of reductively activated peptide gene delivery agents.
AuthorsMcKenzie DL, Kwok KY, Rice KG
JournalJ Biol Chem
PubMed ID10744672
'A new class of peptide gene delivery agents were developed by inserting multiple cysteine residues into short (dp 20) synthetic peptides. Substitution of one to four cysteine residues for lysine residues in Cys-Trp-Lys(18) resulted in low molecular weight DNA condensing peptides that spontaneously oxidize after binding to plasmid DNA to ... More
Biological properties of human prolactin analogs depend not only on global hormone affinity, but also on the relative affinities of both receptor binding sites.
AuthorsKinet S, Bernichtein S, Kelly PA, Martial JA, Goffin V
JournalJ Biol Chem
PubMed ID10473550
'Zinc increases the affinity of human growth hormone (hGH) for the human prolactin receptor (hPRLR) due to the coordination of one zinc ion involving Glu-174(hGH) and His-18(hGH). In contrast, binding of hPRL to the hPRLR is zinc-independent. We engineered in binding site 1 of hPRL a hGH-like zinc coordination site, ... More
Cell-free reconstitution of microautophagic vacuole invagination and vesicle formation.
AuthorsSattler T, Mayer A
JournalJ Cell Biol
PubMed ID11062255
'Many organelles change their shape in the course of the cell cycle or in response to environmental conditions. Lysosomes undergo drastic changes of shape during microautophagocytosis, which include the invagination of their boundary membrane and the subsequent scission of vesicles into the lumen of the organelle. The mechanism driving these ... More
Vav2 is an activator of Cdc42, Rac1, and RhoA.
AuthorsAbe K, Rossman KL, Liu B, Ritola KD, Chiang D, Campbell SL, Burridge K, Der CJ
JournalJ Biol Chem
PubMed ID10744696
'Vav and Vav2 are members of the Dbl family of proteins that act as guanine nucleotide exchange factors (GEFs) for Rho family proteins. Whereas Vav expression is restricted to cells of hematopoietic origin, Vav2 is widely expressed. Although Vav and Vav2 share highly related structural similarities and high sequence identity ... More
The use of ATP bioluminescence as a measure of cell proliferation and cytotoxicity.
'Adenosine triphosphate (ATP) bioluminescence was used to determine whether there was a linear relationship between cultured cell number and measured luminescence using the luciferin-luciferase reaction. In all the cells tested including peripheral blood mononuclear cells (MNC), MOLT-4, HL-60, TF-1, NFS-60 and L-929 cell lines there was a significant correlation as ... More
Inhibition of glucocorticoid-induced apoptosis by the expression of antisense gene of mitochondrial ATPase subunit 6(1).
AuthorsSato M, Matsuki Y, Oguma T, Tsujimoto K, Takayama E, Tadakuma T
JournalFEBS Lett
PubMed ID10922465
'To isolate the apoptosis-linked genes involved in the cell death of thymocytes induced by glucocorticoids, we developed a functional cloning assay. Murine CD4(+)CD8(+) thymic cell line 2-257-20 cells were transfected with cDNA expression libraries obtained from a dexamethasone-resistant cell line. The transfected cells were selected in the presence of dexamethasone, ... More
ATP luminescence-based motility-invasion assay.
Authorsde la Monte SM, Lahousse SA, Carter J, Wands JR
JournalBiotechniques
PubMed ID12139262
'Directional motility and invasion assays are largely based on the use of Boyden chambers or Transwell culture inserts in which porous membranes separate seeded cells from a chemotactic factor supplied in the medium outside the chamber. The major obstaclefor most currently available assays is that they lack a sensitive, easy, ... More
The human immunodeficiency virus-1 Tat protein activates human umbilical vein endothelial cell E-selectin expression via an NF-kappa B-dependent mechanism.
'Human immunodeficiency virus infection is associated with inflammation and endothelial cell activation that cannot be ascribed to direct infection by the virus or to the presence of opportunistic infections. Factors related to the virus itself, to the host and/or to environmental exposures probably account for these observations. The HIV protein ... More
A novel p75TNF receptor isoform mediating NFkappa B activation.
AuthorsSeitz C, Muller P, Krieg RC, Mannel DN, Hehlgans T
JournalJ Biol Chem
PubMed ID11279196
'We report the identification of a novel p75TNF receptor isoform termed icp75TNFR, which is generated by the use of an alternative transcriptional start site within the p75TNFR gene and characterized by regulated intracellular expression. The icp75TNFR protein has an apparent molecular mass of approximately 50 kDa and is recognized by ... More
Determination of sterilization effectiveness by measuring bacterial growth in a biological indicator through firefly luciferase determination of ATP.
AuthorsWebster JJ, Walker BG, Ford SR, Leach FR
JournalJ Biolumin Chemilumin
PubMed ID3213598
'A bioluminescence procedure for measurement of microbial ATP allows a rapid determination of the effectiveness of autoclave sterilization. This determination is achieved faster than detection of acid production in a biological indicator via a pH indicator. Bacterial outgrowth from spores on test strips of the biological indicator was detected by ... More
Bioluminescence and chemiluminescence literature. Luciferase reporter genes--lux and luc. Part 2.
AuthorsHill PJ, Stewart GS, Stanley PE
JournalJ Biolumin Chemilumin
PubMed ID8237470
'The following references encompass a review of recent literature where the in vivo expression of eukaryotic and/or prokaryotic luciferases provide for sensitive reporters of cellular activity. The list is subdivided into prokaryotic and eukaryotic applications. We have included the uses of luciferases in elucidating the control of gene expression or ... More
Optimized use of the firefly luciferase assay as a reporter gene in mammalian cell lines.
AuthorsBrasier AR, Tate JE, Habener JF
JournalBiotechniques
PubMed ID2698191
'The enzymatic activity of firefly luciferase provides a sensitive, rapid means to assay transcriptional activity of regulated activation sequences of DNA when fused to the protein coding sequence of the luciferase gene. We have developed several modifications of the luciferase assay and have characterized certain parameters of the assay, resulting ... More
A microchip for quantitative detection of molecules utilizing luminescent and radioisotope reporter groups.
AuthorsEggers M, Hogan M, Reich RK, Lamture J, Ehrlich D, Hollis M, Kosicki B, Powdrill T, Beattie K, Smith S
JournalBiotechniques
PubMed ID7818906
'Through the marriage of microelectronics and molecular biology, a miniaturized device is presented for ultrasensitive detection of labeled molecules. The novelty of the approach is the direct integration of a charge-coupled device (CCD) and a probe-based assay. Specifically, a CCD detector serves as an active solid support that quantitatively detects ... More
Cytoprotection of human umbilical vein endothelial cells against apoptosis and CTL-mediated lysis provided by caspase-resistant Bcl-2 without alterations in growth or activation responses.
AuthorsZheng L, Dengler TJ, Kluger MS, Madge LA, Schechner JS, Maher SE, Pober JS, Bothwell AL
JournalJ Immunol
PubMed ID10779771
'Graft endothelial cells are primary targets of host CTL-mediated injury in acute allograft rejection. As an in vitro trial of gene therapy to reduce CTL-mediated endothelial injury, we stably transduced early passage HUVEC with a caspase-resistant mutant form (D34A) of the anti-apoptotic gene Bcl-2. Bcl-2 transductants were compared with HUVEC ... More
Clinical and biochemical applications of luciferases and luciferins.
AuthorsKricka LJ
JournalAnal Biochem
PubMed ID3072882
'Recent advances in the analytical applications of bacterial and firefly luciferases and firefly luciferin are reviewed. Luciferases have been used in soluble and immobilized/co-immobilized forms in assays for a variety of enzymes, substrates, and cofactors. The firefly luciferase reaction forms the basis of rapid microbiological tests which have found application ... More
Expression of the gene encoding firefly luciferase in insect cells using a baculovirus vector.
AuthorsHasnain SE, Nakhai B
JournalGene
PubMed ID2205537
'A cDNA encoding the firefly luciferase [Photinus luciferin: oxygen 4-oxidoreductase (decarboxylating, ATP-hydrolyzing), EC 1.13.12.7] was cloned downstream from the polyhedrin gene promoter of Autographa californica nuclear polyhedrosis virus and expressed in Spodoptera frugiperda clone-9 cells. Synthesis of luciferase (Luc) was accurately measured in insect cells growing in a 96-well plate, ... More
Preparation of a genetically fused protein A/luciferase conjugate for use in bioluminescent immunoassays.
AuthorsLindbladh C, Mosbach K, Bülow L
JournalJ Immunol Methods
PubMed ID2013697
'The genes encoding staphylococcal protein A and bacterial luciferase (Vibrio harveyi) were fused in-frame in order to obtain a general marker enzyme for bioluminescent immunoassays. Two constructs were made where protein A was ligated to the first and the 12th amino acid residue, respectively, of the N terminus of the ... More
Functional cloning of the proto-oncogene brain factor-1 (BF-1) as a Smad-binding antagonist of transforming growth factor-beta signaling.
AuthorsRodriguez C, Huang LJ, Son JK, McKee A, Xiao Z, Lodish HF
JournalJ Biol Chem
PubMed ID11387330
'Using the plasminogen activator inhibitor (PAI) promoter to drive the expression of a reporter gene (mouse CD2), we devised a system to clone negative regulators of the transforming growth factor-beta (TGF-beta) signaling pathway. We infected a TGF-beta-responsive cell line (MvLu1) with a retroviral cDNA library, selecting by fluorescence-activated cell sorter ... More
A review of bioluminescent ATP techniques in rapid microbiology.
AuthorsStanley PE
JournalJ Biolumin Chemilumin
PubMed ID2678922
'Use of firefly luciferase to assay adenosine triphosphate (ATP) extracted from microorganisms provides an easy means to enumerate microbes within minutes. The small amount of light produced is proportional to ATP and thus microbial number. The average bacterium contains around 10(-15) g ATP per cell. Present reagents permit detection of ... More
Firefly luciferase, synthesized to very high levels in caterpillars infected with a recombinant baculovirus, can also be used as an efficient reporter enzyme in vivo.
AuthorsJha PK, Nakhai B, Sridhar P, Talwar GP, Hasnain SE
JournalFEBS Lett
PubMed ID2253776
'Trichoplusia ni and Spodoptera littoralis larvae were infected with a recombinant AcNPV, having the viral polyhedrin gene replaced with the cDNA encoding firefly luciferase. Both S. littoralis and T. ni synthesized very high levels of luciferase representing greater than or equal to 25% and greater than or equal to 15%, ... More
Low intracellular zinc impairs the translocation of activated NF-kappa B to the nuclei in human neuroblastoma IMR-32 cells.
Authors Mackenzie Gerardo G; Zago M Paola; Keen Carl L; Oteiza Patricia I;
JournalJ Biol Chem
PubMed ID12089148
'In the current work, we studied how variations in extracellular zinc concentrations modulate different steps involved in nuclear factor kappaB (NF-kappaB) activation in human neuroblastoma IMR-32 cells. Cells were incubated in media containing varying concentrations of zinc (1.5, 5, 15, and 50 microm). Within 3 h, the intracellular zinc content ... More
A new type of ultrasensitive bioluminogenic enzyme substrates. I. Enzyme substrates with D-luciferin as leaving group.
AuthorsMiska W, Geiger R
JournalBiol Chem Hoppe Seyler
PubMed ID3166746
'Derivatives of D-luciferin, D-luciferin methyl ester, D-luciferin O-sulfate, D-luciferin O-phosphate, D-luciferyl-L-N alpha-arginine and D-luciferyl-L-phenylalanine were used as highly sensitive substrates for carboxylic esterase, arylsulfatase, alkaline phosphatase and carboxypeptidases A, B and N. Enzymatic cleavage of the compounds by enzymes leading to the release of D-luciferin was demonstrated. Kinetic constants have ... More
Cell biology. NO helps make fireflies flash.
AuthorsPennisi E
JournalScience
PubMed ID11431541
Normalization of luciferase reporter assays under conditions that alter internal controls.
AuthorsSims RJ, Liss AS, Gottlieb PD
JournalBiotechniques
PubMed ID12765018
Membrane targeting of firefly luciferase: a new bioluminescent reporter gene.
AuthorsBadia E, Duchesne MJ, Pons M, Nicolas JC
JournalAnal Biochem
PubMed ID8203765
Research at the Interface between Chemistry and Virology: Development of a Molecular Flashlight.
AuthorsKraus GA, Zhang W, Fehr MJ, Petrich JW, Wannemuehler Y, Carpenter S
JournalChem Rev
PubMed ID11848763
Reporter genes and transient assays for plants.
AuthorsMatthews BF, Saunders JA, Gebhardt JS, Lin JJ, Koehler SM
JournalMethods Mol Biol
PubMed ID8528416
Reporter genes.
AuthorsSuter-Crazzolara C, Klemm M, Reiss B
JournalMethods Cell Biol
PubMed ID8531814
Chemiluminescent and bioluminescent reporter gene assays.
AuthorsBronstein I, Fortin J, Stanley PE, Stewart GS, Kricka LJ
JournalAnal Biochem
PubMed ID8080073
Bioluminescent continuous-flow assay of adenosine 5'-triphosphate using firefly luciferase immobilized on nylon tubes.
AuthorsCarrea G, Bovara R, Mazzola G, Girotti S, Roda A, Ghini S
JournalAnal Chem
PubMed ID3963392
Cloning firefly luciferase.
Authorsde Wet JR, Wood KV, Helinski DR, DeLuca M
JournalMethods Enzymol
PubMed ID3547031
Luciferase reporter gene cassettes for plant gene expression studies.
AuthorsRiggs CD, Chrispeels MJ
JournalNucleic Acids Res
PubMed ID3671076
Use of the liquid scintillation spectrometer for determining adenosine triphosphate by the luciferase enzyme.
AuthorsStanley PE, Williams SG
JournalAnal Biochem
PubMed ID4307203
Extraction of adenosine triphosphate from microbial and somatic cells.
AuthorsStanley PE
JournalMethods Enzymol
PubMed ID3821533
Spatial imaging of luciferase gene expression in transgenic fish.
AuthorsTamiya E, Sugiyama T, Masaki K, Hirose A, Okoshi T, Karube I
JournalNucleic Acids Res
PubMed ID2315029
lux genes and the applications of bacterial bioluminescence.
AuthorsStewart GS, Williams P
JournalJ Gen Microbiol
PubMed ID1512559
Transient expression analysis in plants using firefly luciferase reporter gene.