Showing papers in "Nature in 1995"

Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B.

Abstract: Glycogen synthase kinase-3 (GSK3) is implicated in the regulation of several physiological processes, including the control of glycogen and protein synthesis by insulin, modulation of the transcription factors AP-1 and CREB, the specification of cell fate in Drosophila and dorsoventral patterning in Xenopus embryos. GSK3 is inhibited by serine phosphorylation in response to insulin or growth factors and in vitro by either MAP kinase-activated protein (MAPKAP) kinase-1 (also known as p90rsk) or p70 ribosomal S6 kinase (p70S6k). Here we show, however, that agents which prevent the activation of both MAPKAP kinase-1 and p70S6k by insulin in vivo do not block the phosphorylation and inhibition of GSK3. Another insulin-stimulated protein kinase inactivates GSK3 under these conditions, and we demonstrate that it is the product of the proto-oncogene protein kinase B (PKB, also known as Akt/RAC). Like the inhibition of GSK3 (refs 10, 14), the activation of PKB is prevented by inhibitors of phosphatidylinositol (PI) 3-kinase.

Rapid turnover of plasma virions and CD4 lymphocytes in HIV-1 infection

Abstract: Treatment of infected patients with ABT-538, an inhibitor of the protease of human immunodeficiency virus type 1 (HIV-1), causes plasma HIV-1 levels to decrease exponentially (mean half-life, 2.1 +/- 0.4 days) and CD4 lymphocyte counts to rise substantially. Minimum estimates of HIV-1 production and clearance and of CD4 lymphocyte turnover indicate that replication of HIV-1 in vivo is continuous and highly productive, driving the rapid turnover of CD4 lymphocytes.

Cloning of a gene bearing missense mutations in early-onset familial Alzheimer's disease

Abstract: Some cases of Alzheimer's disease are inherited as an autosomal dominant trait. Genetic linkage studies have mapped a locus (AD3) associated with susceptibility to a very aggressive form of Alzheimer's disease to chromosome 14q24.3. We have defined a minimal cosegregating region containing the AD3 gene, and isolated at least 19 different transcripts encoded within this region. One of these transcripts (S182) corresponds to a novel gene whose product is predicted to contain multiple transmembrane domains and resembles an integral membrane protein. Five different missense mutations have been found that cosegregate with early-onset familial Alzheimer's disease. Because these changes occurred in conserved domains of this gene, and are not present in normal controls, they are likely to be causative of AD3.

Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis

Abstract: The protease responsible for the cleavage of poly(ADP-ribose) polymerase and necessary for apoptosis has been purified and characterized. This enzyme, named apopain, is composed of two subunits of relative molecular mass (M(r)) 17K and 12K that are derived from a common proenzyme identified as CPP32. This proenzyme is related to interleukin-1 beta-converting enzyme (ICE) and CED-3, the product of a gene required for programmed cell death in Caenorhabditis elegans. A potent peptide aldehyde inhibitor has been developed and shown to prevent apoptotic events in vitro, suggesting that apopain/CPP32 is important for the initiation of apoptotic cell death.

Failure of blood-island formation and vasculogenesis in Flk-1-deficient mice.

Abstract: The receptor tyrosine kinase Flk-1 (ref. 1) is believed to play a pivotal role in endothelial development. Expression of the Flk-1 receptor is restricted to endothelial cells and their embryonic precursors, and is complementary to that of its ligand, vascular endothelial growth factor (VEGF), which is an endothelial-specific mitogen. Highest levels of flk-1 expression are observed during embryonic vasculogenesis and angiogenesis, and during pathological processes associated with neovascularization, such as tumour angiogenesis. Because flk-1 expression can be detected in presumptive mesodermal yolk-sac blood-island progenitors as early as 7.0 days postcoitum, Flk-1 may mark the putative common embryonic endothelial and haematopoietic precursor, the haemangioblast, and thus may also be involved in early haematopoiesis. Here we report the generation of mice deficient in Flk-1 by disruption of the gene using homologous recombination in embryonic stem (ES) cells. Embryos homozygous for this mutation die in utero between 8.5 and 9.5 days post-coitum, as a result of an early defect in the development of haematopoietic and endothelial cells. Yolk-sac blood islands were absent at 7.5 days, organized blood vessels could not be observed in the embryo or yolk sac at any stage, and haematopoietic progenitors were severely reduced. These results indicate that Flk-1 is essential for yolk-sac blood-island formation and vasculogenesis in the mouse embryo.

A Jupiter-Mass Companion to a Solar-Type Star

Abstract: The presence of a Jupiter-mass companion to the star 51 Pegasi is inferred from observations of periodic variations in the star's radial velocity. The companion lies only about eight million kilometres from the star, which would be well inside the orbit of Mercury in our Solar System. This object might be a gas-giant planet that has migrated to this location through orbital evolution, or from the radiative stripping of a brown dwarf.

Cpg motifs in bacterial dna trigger direct b-cell activation

Abstract: Unmethylated CpG dinucleotides are more frequent in the genomes of bacteria and viruses than of vertebrates. We report here that bacterial DNA and synthetic oligodeoxynucleotides containing unmethylated CpG dinucleotides induce murine B cells to proliferate and secrete immunoglobulin in vitro and in vivo. This activation is enhanced by simultaneous signals delivered through the antigen receptor. Optimal B-cell activation requires a DNA motif in which an unmethylated CpG dinucleotide is flanked by two 5' purines and two 3' pyrimidines. Oligodeoxynucleotides containing this CpG motif induce more than 95% of all spleen B cells to enter the cell cycle. These data suggest a possible evolutionary link between immune defence based on the recognition of microbial DNA and the phenomenon of 'CpG suppression' in vertebrates. The potent immune activation by CpG oligonucleotides has implications for the design and interpretation of studies using 'antisense' oligonucleotides and points to possible new applications as adjuvants.

Identification of the breast cancer susceptibility gene BRCA2

Abstract: IN Western Europe and the United States approximately 1 in 12 women develop breast cancer. A small proportion of breast cancer cases, in particular those arising at a young age, are attributable to a highly penetrant, autosomal dominant predisposition to the disease. The breast cancer susceptibility gene, BRCA2, was recently localized to chromosome 13q12-q13. Here we report the identification of a gene in which we have detected six different germline mutations in breast cancer families that are likely to be due to BRCA2. Each mutation causes serious disruption to the open reading frame of the transcriptional unit. The results indicate that this is the BRCA2 gene.

Principles of CDK regulation

Abstract: As key regulators of the cell cycle, the cyclin-dependent kinases must be tightly regulated by extra- and intracellular signals. The activity of cyclin-dependent kinases is controlled by four highly conserved biochemical mechanisms, forming a web of regulatory pathways unmatched in its elegance and intricacy.

Viral dynamics in human immunodeficiency virus type 1 infection

Abstract: The dynamics of HIV-1 replication in vivo are largely unknown yet they are critical to our understanding of disease pathogenesis. Experimental drugs that are potent inhibitors of viral replication can be used to show that the composite lifespan of plasma virus and virus-producing cells is remarkably short (half-life approximately 2 days). Almost complete replacement of wild-type virus in plasma by drug-resistant variants occurs after fourteen days, indicating that HIV-1 viraemia is sustained primarily by a dynamic process involving continuous rounds of de novo virus infection and replication and rapid cell turnover.

Efficient photodiodes from interpenetrating polymer networks

Abstract: THE photovoltaic effect involves the production of electrons and holes in a semiconductor device under illumination, and their subsequent collection at opposite electrodes. In many inorganic semiconductors, photon absorption produces free electrons and holes directly1. But in molecular semiconductors, absorption creates electrona¤-hole pairs (excitons) which are bound at room temperature2, so that charge collection requires their dissociation. Exciton dissociation is known to be efficient at interfaces between materials with different electron affinities and ionization potentials, where the electron is accepted by the material with larger electron affinity and the hole by the material with lower ionization potential3. A two-layer diode structure can thus be used, in which excitons generated in either layer diffuse towards the interface between the layers. However, the exciton diffusion range is typically at least a factor of 10 smaller than the optical absorption depth, thus limiting the efficiency of charge collection3. Here we show that the interpenetrating network formed from a phase-segregated mixture of two semiconducting polymers provides both the spatially distributed interfaces necessary for efficient charge photo-generation, and the means for separately collecting the electrons and holes. Devices using thin films of these polymer mixtures show promise for large-area photodetectors.

Economics Of Climate Change

Abstract: Global climate change poses a threat to the well-being of humans and other living things through impacts on ecosystem functioning, biodiversity, capital productivity, and human health. Climate change economics attends to this issue by offering theoretical insights and empirical findings relevant to the design of policies to reduce, avoid, or adapt to climate change. This economic analysis has yielded new estimates of mitigation benefits, improved understanding of costs in the presence of various market distortions or imperfections, better tools for making policy choices under uncertainty, and alternate mechanisms for allowing flexibility in policy responses. These contributions have influenced the formulation and implementation of a range of climate change policies at the domestic and international levels.

Role of the Flt-1 receptor tyrosine kinase in regulating the assembly of vascular endothelium

Abstract: The vascular endothelial growth factor (VEGF) and its high-affinity binding receptors, the tyrosine kinases Flt-1 and Flk-1, are thought to be important for the development of embryonic vasculature. Here we report that Flt-1 is essential for the organization of embryonic vasculature, but is not essential for endothelial cell differentiation. Mouse embryos homozygous for a targeted mutation in the flt-1 locus, flt-1lcz, formed endothelial cells in both embryonic and extra-embryonic regions, but assembled these cells into abnormal vascular channels and died in utero at mid-somite stages. At earlier stages, the blood islands of flt-1lcz homozygotes were abnormal, with angioblasts in the interior as well as on the periphery. We suggest that the Flt-1 signalling pathway may regulate normal endothelial cell-cell or cell-matrix interactions during vascular development.

Why gold is the noblest of all the metals

Abstract: THE unique role that gold plays in society is to a large extent related to the fact that it is the most noble of all metals: it is the least reactive metal towards atoms or molecules at the interface with a gas or a liquid. The inertness of gold does not reflect a general inability to form chemical bonds, however—gold forms very stable alloys with many other metals. To understand the nobleness of gold, we have studied a simple surface reaction, the dissociation of H2 on the surface of gold and of three other metals (copper, nickel and platinum) that lie close to it in the periodic table. We present self-consistent density-functional calculations of the activation barriers and chemisorption energies which clearly illustrate that nobleness is related to two factors: the degree of filling of the antibonding states on adsorption, and the degree of orbital overlap with the adsorbate. These two factors, which determine both the strength of the adsorbate-metal interaction and the energy barrier for dissociation, operate together to the maxima] detriment of adsorbate binding and subsequent reactivity on gold.

Alzheimer-type neuropathology in transgenic mice overexpressing V717F beta-amyloid precursor protein.

Abstract: Alzheimer's disease (AD) is the most common cause of progressive intellectual failure in aged humans. AD brains contain numerous amyloid plaques surrounded by dystrophic neurites, and show profound synaptic loss, neurofibrillary tangle formation and gliosis. The amyloid plaques are composed of amyloid beta-peptide (A beta), a 40-42-amino-acid fragment of the beta-amyloid precursor protein (APP). A primary pathogenic role for APP/A beta is suggested by missense mutations in APP that are tightly linked to autosomal dominant forms of AD. A major obstacle to elucidating and treating AD has been the lack of an animal model. Animals transgenic for APP have previously failed to show extensive AD-type neuropathology, but we now report the production of transgenic mice that express high levels of human mutant APP (with valine at residue 717 substituted by phenylalanine) and which progressively develop many of the pathological hallmarks of AD, including numerous extracellular thioflavin S-positive A beta deposits, neuritic plaques, synaptic loss, astrocytosis and microgliosis. These mice support a primary role for APP/A beta in the genesis of AD and could provide a preclinical model for testing therapeutic drugs.

Crystal structure of an integral membrane light-harvesting complex from photosynthetic bacteria

Abstract: The crystal structure of the light-harvesting antenna complex (LH2) from Rhodopseudomonas acidophila strain 10050 shows that the active assembly consists of two concentric cylinders of helical protein subunits which enclose the pigment molecules Eighteen bacteriochlorophyll a molecules sandwiched between the helices form a continuous overlapping ring, and a further nine are positioned between the outer helices with the bacteriochlorin rings perpendicular to the transmembrane helix axis There is an elegant intertwining of the bacteriochlorophyll phytol chains with carotenoid, which spans the complex

Cloning of a new cytokine that induces IFN-gamma production by T cells.

Abstract: The mechanism underlying the differentiation of CD4+ T cells into functionally distinct subsets (Th1 and Th2) is incompletely understood, and hitherto unidentified cytokines may be required for the functional maturation of these cells. Here we report the cloning of a recently identified IFN-gamma-inducing factor (IGIF) that augments natural killer (NK) activity in spleen cells. The gene encodes a precursor protein of 192 amino acids and a mature protein of 157 amino acids, which have no obvious similarities to any peptide in the databases. Messenger RNAs for IGIF and interleukin-12 (IL-12) are readily detected in Kupffer cells and activated macrophages. Recombinant IGIF induces IFN-gamma more potently than does IL-12, apparently through a separate pathway. Administration of anti-IGIF antibodies prevents liver damage in mice inoculated with Propionibacterium acnes and challenged with lipopolysaccharide, which induces toxic shock. IGIF may be involved in the development of Th1 cells and also in mechanisms of tissue injury in inflammatory reactions.

Evidence for stripe correlations of spins and holes in copper oxide superconductors

Abstract: ONE of the long-standing mysteries associated with the high-temperature copper oxide superconductors concerns the anomalous suppression1 of superconductivity in La2-xBaxCuO4 (and certain related compounds) when the hole concentration x is near . Here we examine the possibility that this effect is related to dynamical two-dimensional spin correlations, incommensurate with the crystal lattice, that have been observed in La2-xSrxCuO4 by neutron scattering2–4. A possible explanation for the incommensurability involves a coupled, dynamical modulation of spin and charge in which antiferromagnetic 'stripes' of copper spins are separated by periodically spaced domain walls to which the holes segregate5–9. An ordered stripe phase of this type has recently been observed in hole-doped La2NiO4 (refs 10–12). We present evidence from neutron diffraction that in the copper oxide material La1.6-xNd0.4SrxCuO4, with x = 0.12, a static analogue of the dynamical stripe phase is present, and is associated with an anomalous suppression of superconductivity13,14. Our results thus provide an explanation of the ' ' conundrum, and also support the suggestion15 that spatial modulations of spin and charge density are related to superconductivity in the copper oxides.

Protein modules and signalling networks

Abstract: Communication between cells assumes particular importance in multicellular organisms. The growth, migration and differentiation of cells in the embryo, and their organization into specific tissues, depend on signals transmitted from one cell to another. In the adult, cell signalling orchestrates normal cellular behaviour and responses to wounding and infection. The consequences of breakdowns in this signalling underlie cancer, diabetes and disorders of the immune and cardiovascular systems. Conserved protein domains that act as key regulatory participants in many of these different signalling pathways are highlighted.

Fatigue-free ferroelectric capacitors with platinum electrodes

Abstract: A SIGNIFICANT fraction of the computer memory industry is at present involved in the manufacture of non-volatile memory devices1—that is, devices which retain information when power is interrupted. For such applications (and also for volatile memories), the use of capacitors constructed from ferroelectric thin films has stimulated much interest1. In such structures, information is stored in the polarization state of the ferroelectric material itself, which should in principle lead to lower power requirements, faster access time and potentially lower cost1. But the use of ferroelectrics is not without problems; the memories constructed to date have generally suffered from poor retention of stored information and degradation of performance ('fatigue') with use1–3. Here we describe the preparation and characterization of thin-film capacitors using ferroelectric materials from a large family of layered perovskite oxides, exemplified by SrBi2Ta2O9, SrBi2NbTaO9 and SrBi4Ta4O15. The structural flexibility of these materials allows their properties to be tailored so that many of the problems associated with previous ferroelectric memories are avoided. In particular, our capacitors do not show significant fatigue after 1012 switching cycles, and they exhibit good retention characteristics and low leakage currents even with films less than 100 nm thick.

Protein kinase B (c-Akt) in phosphatidylinositol-3-OH kinase signal transduction

Abstract: A serine/threonine kinase, named protein kinase B (PKB) for its sequence homology to both protein kinase A and C, has previously been isolated. PKB, which is identical to the kinase Rac, was later found to be the cellular homologue of the transforming v-Akt. Here we show that PKB is activated by stimuli such as insulin, platelet-derived growth factor (PDGF), epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF). Activation of PKB was inhibited by the phosphatidylinositol-3-OH kinase (PI(3)K) inhibitor wortmannin and by coexpression of a dominant-negative mutant of PI(3)K. PDGF receptor mutants that lack detectable associated PI(3)K activity also fail to induce PKB activation, PKB kinase activity is correlated with phosphorylation of PKB on serine. Finally, we show that a constructed Gag-PKB fusion protein, homologous to the v-akt oncogene, displays significantly increased ligand-independent kinase activity. Furthermore, this activity is sufficient to activate the p70 S6-kinase (p70S6K). These results suggest a role for PKB in PI(3)K-mediated signal transduction.

Selective binding and removal of guests in a microporous metal–organic framework

Abstract: MICROPOROUS inorganic materials such as zeolites find widespread application in heterogeneous catalysis, adsorption and ion-exchange processes. The rigidity and stability of such frameworks allow for shape- and size-selective inclusion of organic molecules and ions1–4. Analogous microporous structures based on organic building blocks have the potential for more precise rational design, through control of the shape, size and functionalization of the pores5–8. Here we report the synthesis of a metal–organic framework designed to bind aromatic guest molecules selectively. The basic building block is a symmetric organic molecule, which binds metal ions9,10 to form layers of the metal–organic compound alternating with layers whose composition is determined by the functionalization of the starting molecules. The layers create channels in which guest aromatic molecules may be selectively bound. We show that the crystal lattice thus formed is thermally stable up to 350 °C, even after removal of included guest molecules, and that the inclusions can be selectively readsorbed.

Transcriptional co-repressor that interacts with nuclear hormone receptors

Abstract: In accordance with the present invention, there are provided novel receptor interacting factors, referred to herein as “SMRT”, i.e., a silencing mediator (co-repressor) for retinoic acid receptor (RAR) and thyroid hormone receptor (TR). SMRT is a novel protein whose association with RAR and TR both in solution and on DNA response elements is destabilized by ligand. The interaction of SMRT with mutant receptors correlates with the transcriptional silencing activities of receptors. In vivo, SMRT functions as a potent co-repressor. A GAL4 DNA binding domain (DBD) fusion of SMRT behaves as a frank repressor of a GAL4-dependent reporter. Together, these data identify a novel class of cofactor which is believed to represent an important mediator of hormone action.

Ligand-independent repression by the thyroid hormone receptor mediated by a nuclear receptor co-repressor

Abstract: Thyroid-hormone and retinoic-acid receptors exert their regulatory functions by acting as both activators and repressors of gene expression. A nuclear receptor co-repressor (N-CoR) of relative molecular mass 270K has been identified which mediates ligand-independent inhibition of gene transcription by these receptors, suggesting that the molecular mechanisms of repression by thyroid-hormone and retinoic-acid receptors are analogous to the co-repressor-dependent transcriptional inhibitory mechanisms of yeast and Drosophila.

Familial Alzheimer's disease in kindreds with missense mutations in a gene on chromosome 1 related to the Alzheimer's disease type 3 gene

Abstract: We report the cloning of a novel gene (E5-1) encoded on chromosome 1 which has substantial nucleotide and amino-acid sequence similarity to the S182 gene on chromosome 14q24.3. Mutations, including three new missense mutations in the S182 gene, are associated with the AD3 subtype of early-onset familial Alzheimer's disease (AD). Both the E5-1 and the S182 proteins are predicted to be integral membrane proteins with seven membrane-spanning domains, and a large exposed loop between the sixth and seventh transmembrane domains. Analysis of the nucleotide sequence of the open reading frame (ORF) of the E5-1 gene led to the discovery of two missense substitutions at conserved amino-acid residues in affected members of pedigrees with a form of familial AD that has a later age of onset than the AD3 subtype (50-70 years versus 30-60 years for AD3). These observations imply that the E5-1 gene on chromosome 1 and the S182 gene on chromosome 14q24.3 are members of a family of genes (presenilins) with related functions, and indicates that mutations in conserved residues of E5-1 could also play a role in the genesis of AD. Our results also indicate that still other AD susceptibility genes exist.

Hypertension in mice lacking the gene for endothelial nitric oxide synthase

Abstract: Nitric oxide (NO), a potent vasodilator produced by endothelial cells, is thought to be the endothelium-dependent relaxing factor (EDRF) which mediates vascular relaxation in response to acetylcholine, bradykinin and substance P in many vascular beds. NO has been implicated in the regulation of blood pressure and regional blood flow, and also affects vascular smooth-muscle proliferation and inhibits platelet aggregation and leukocyte adhesion. Abnormalities in endothelial production of NO occur in atherosclerosis, diabetes and hypertension. Pharmacological blockade of NO production with arginine analogues such as L-nitroarginine (L-NA) or L-N-arginine methyl ester affects multiple isoforms of nitric oxide synthase (NOS), and so cannot distinguish their physiological roles. To study the role of endothelial NOS (eNOS) in vascular function, we disrupted the gene encoding eNOS in mice. Endothelium-derived relaxing factor activity, as assayed by acetylcholine-induced relaxation, is absent, and the eNOS mutant mice are hypertensive. Thus eNOS mediates basal vasodilation. Responses to NOS blockade in the mutant mice suggest that non-endothelial isoforms of NOS may be involved in maintaining blood pressure.

The synaptic vesicle cycle: a cascade of protein–protein interactions

Abstract: The synaptic vesicle cycle at the nerve terminal consists of vesicle exocytosis with neurotransmitter release, endocytosis of empty vesicles, and regeneration of fresh vesicles. Of all cellular transport pathways, the synaptic vesicle cycle is the fastest and the most tightly regulated. A convergence of results now allows formulation of molecular models for key steps of the cycle. These developments may form the basis for a mechanistic understanding of higher neural function.

Structure at 2.8 A resolution of cytochrome c oxidase from Paracoccus denitrificans.

Abstract: The crystal structure at 2.8 A resolution of the four protein subunits containing cytochrome c oxidase from the soil bacterium Paracoccus denitrificans, complexed with an antibody Fv fragment, is described. Subunit I contains 12 membrane-spanning, primarily helical segments and binds haem a and the haem a3-copper B binuclear centre where molecular oxygen is reduced to water. Two proton transfer pathways, one for protons consumed in water formation and one for 'proton pumping', could be identified. Mechanisms for proton pumping are discussed.

Isolation and structure of the endogenous agonist of opioid receptor-like ORL1 receptor.

Abstract: The ORL1 receptor, an orphan receptor whose human and murine complementary DNAs have recently been characterized, structurally resembles opioid receptors and is negatively coupled with adenylate cyclase. ORL1 transcripts are particularly abundant in the central nervous system. Here we report the isolation, on the basis of its ability to inhibit the cyclase in a stable recombinant CHO(ORL1+) cell line, of a neuropeptide that resembles dynorphin A9 and whose amino acid sequence is Phe-Gly-Gly-Phe-Thr-Gly-Ala-Arg-Lys-Ser-Ala-Arg-Lys-Leu-Ala-Asn-Gln. The rat-brain cDNA encodes the peptide flanked by Lys-Arg proteolytic cleavage motifs. The synthetic heptadecapeptide potently inhibits adenylate cyclase in CHO(ORL1+) cells in culture and induces hyperalgesia when administered intracerebroventricularly to mice. Taken together, these data indicate that the newly discovered heptadecapeptide is an endogenous agonist of the ORL1 receptor and that it may be endowed with pro-nociceptive properties.

Reduced cell motility and enhanced focal adhesion contact formation in cells from FAK-deficient mice

Abstract: The intracellular protein tyrosine kinase FAK (focal adhesion kinase) was originally identified gy its high level of tyrosine phosphorylation in v-src-transformed cells. FAK is also highly phosphorylated during early development. In cultured cells it is localized to focal adhesion contacts and becomes phosphorylated and activated in response to integrin-mediated binding of cells to the extracellular matrix, suggesting an important role in cell adhesion and/or migration. We have generated FAK-deficient mice by gene targeting to examine the role of FAK during development. Mutant embryos displayed a general defect of mesoderm development, and cells from these embryos had reduced mobility in vitro. Surprisingly, the number of focal adhesions was increased in FAK-deficient cells, suggesting that FAK may be involved in the turnover of focal adhesion contacts during cell migration.