cDNA cloning and expression of a hamster α-thrombin receptor coupled to Ca2+ mobilization

AbstractThe serine protease α-thrombin (thrombin) potently stimulates G-protein-coupled signaling pathways and DNA synthesis in CCL39 hamster lung fibroblasts. To clone a thrombin receptor cDNA, selective amplification of mRNA sequences displaying homology to the transmembrane domains of G-protein-coupled receptor genes was performed by polymerase chain reaction. Using reverse transcribed poly(A)+ RNA from CCL39 cells and degenerate primers corresponding to conserved regions of several phospholipase C-coupled receptors, three novel putative receptor sequences were identified. One corresponds to an mRNA transcript of 3.4 kb in CCL39 cells and a relatively abundant cDNA. Microinjection of RNA transcribed in vitro from this cDNA in Xenopus oocytes leads to the expression of a functional thrombin receptor. The hamster thrombin receptor consists of 427 amino acid residues with 8 hydrophobic domains, including one at the extreme N-terminus that is likely to represent a signal peptide. A thrombin consensus cleavage site is present in the N-terminal extracellular region of the receptor sequence followed by a negatively charged cluster of residues present in a number of proteins that interact with the anion-binding exosite of thrombin

HIV-1 drug-resistance patterns among patients on failing treatment in a large number of European countries

Background: Information about patterns of HIV-1 drug resistance among treatment-exposed patients is crucial for the development of novel effective drugs. Currently no system exists that monitors patterns of resistance in patients failing therapy. Methods: The study included 1,988 HIV-1 sequences from patients experiencing therapy failure collected between 2000 and 2004 in 15 European countries. Genotypic resistance was interpreted using the ANRS algorithm. Phenotypic resistance was predicted using the Virco geno- to phenotype system. Results: 80.7% of the sequences included at least one drug-resistance mutation. Mutations were found for NRTIs (73.5%), NNRTIs (48.5%), and protease inhibitors (35.8%). Ninety percent of sequences with genotypic resistance harbored M184V, M41L, K103N, D67N, and/or T215Y. Among NRTIs, resistance was most frequently predicted for lamivudine. About half of all sequences had reduced susceptibility for NNRTIs. Resistance to most boosted protease inhibitors was found in &lt; 25%. No sequence had resistance to all currently available drugs. Conclusion: Levels of resistance among patients with therapy failure were high. The patterns of resistance reflect resistance to drugs available for a longer time. Fully suppressive regimens can be designed even for the most mutated HIV because boosted protease inhibitors have remained active against most circulating viruses and new drug classes have become available.</p

CITY AUTOMATED TRANSPORT SYSTEM (CATS): THE LEGACY OF AN INNOVATIVE EUROPEAN PROJECT

CATS is a collaborative European project promoting driverless vehicles that ended in December 2014. This contribution explains how the project evolved, including the handling of unexpected events and concentrating on lessons learned. The constructor and vehicle had to be changed for economic reasons in the middle of the project timeline. A second constructor went bankrupt, although access to his vehicles could be secured. For security and legal reasons, part of the final demonstration was relocated at short notice to the EPFL campus in Lausanne, Switzerland, where around 1600 people were transported during 16 days of vehicle operation. Reactions to the driverless vehicle concept were overwhelmingly positive. Implications for the acceptability of driverless vehicles in Europe and elsewhere are discussed

Etude génétique et biochimique de la régulation de la transcription

Doctorat en Sciencesinfo:eu-repo/semantics/nonPublishe

Aspects génétiques et biochimiques de la régulation de la transcription chez la bactérie E. coli et ses phages

SCOPUS: re.jinfo:eu-repo/semantics/publishe

Etude génétique et biochimique de la régulation de la transcription

Doctorat en Sciencesinfo:eu-repo/semantics/nonPublishe

Differential action of trypsin on the β′ subunit of DNA-dependent RNA polymerase from E. coli

SCOPUS: ar.jinfo:eu-repo/semantics/publishe

The firA gene, a locus involved in the expression of rifampicin resistance in Escherichia coli - II. Characterisation of bacterial proteins coded by λfirA transducing phages

The firA gene probably codes for an essential component of the transcription machinery in E. coli. Bacterial proteins coded by λfirA transducing phages have been examined after infection of a UV-irradiated λ lysogen, and 2 major fir-specific proteins have been characterized. The larger protein has a molecular weight of 27,000 daltons. The smaller protein, of 17,000 daltons, is produced in a considerable molar excess over the larger protein, is basic and binds strongly to DNA-, to DEAE-and to phospho-cellulose. This protein is clearly visible upon 2-dimensional gel electrophoresis of unfractionated E. coli protein, showing that it is present in the cell in large quantities. Evidence is presented to suggest that this protein may be identical to the Kappa factor of Schäfer and Zillig (1973). © 1977 Springer-Verlag.SCOPUS: ar.jinfo:eu-repo/semantics/publishe