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J Raimond

Publications and source records attributed to J Raimond.

14 recordsLinked to original sources

Identification of an internal gene to the human Galectin-3 gene with two different overlapping reading frames that do not encode Galectin-3.

We previously reported that alternative transcripts were initiated within the second intron of the human Galectin-3 gene (LGALS3). We now demonstrate that these transcripts arise from an internal gene embedded within LGALS3 and named galig (Galectin-3 internal gene). Tissue-specific expression of galig was assayed by screening of several human tissues. Contrary to LGALS3, galig appears to be tightly regulated and principally activated in leukocytes from peripheral blood. Cloning and characterization of galig transcripts revealed that they contain two out-of-frame overlapping open-reading frames (ORFs). Transfection of expression vectors encoding enhanced green fluorescent protein (EGFP) chimeras indicated that both ORFs could be translated in proteins unrelated to Galectin-3. The ORF1 polypeptide targets EGFP to cytosol and nucleus whereas ORF2 targets EGFP to mitochondria. These results revealed the exceptional genetic organization of the LGALS3 locus.

Amino Acid Sequence↗

Gene transfer by DNA/glycosylated polylysine complexes into human blood monocyte-derived macrophages.

Macrophages are putative target cells for expressing an exogenous gene with therapeutical effects. Knowing that macrophages express membrane lectins mediating endocytosis of their ligands, DNA/glycosylated polylysine complexes were used to transfect human blood monocyte-derived macrophages. Monocytes from human peripheral blood were matured in culture for 7 days to differentiate into macrophage-like cells in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF). Adherent cells, which displayed characteristic macrophage markers, CD 14, CD 11b, HLA-DR, and HLA-ABC antigens and mannose receptor, were transfected by DNA/glycosylated polylysine complexes in the presence of chloroquine. The luciferase reporter gene expression was maximal 24 hr after transfection with a DNA/mannosylated polylysine complex and by using plasmids in which the promoters (either the long terminal repeat of the human immunodeficiency virus or the human cytomegalovirus) drove the luciferase gene expression. Luciferase gene expression was lower when the promoter was the early region of the large T antigen of SV40 virus. Transfection mediated by DNA/mannosylated polylysine complexes was much more efficient than with DEAE-dextran or lipofectin. The possibility of transferring and expressing an exogenous gene into macrophage-like cells by using a nonimmunogenic synthetic vector as a DNA carrier opens new ways to develop nonviral gene therapy strategies.

Cell Count↗

The second intron of the human galectin-3 gene has a strong promoter activity down-regulated by p53.

Galectin-3 is a galactose-specific lectin which has been shown to be involved in several biological functions such as cell growth regulation, cell aggregation and cell differentiation. The partial cloning of the human genomic sequences reveals the presence of a 651 bp intron, 18 bp downstream of the translation initiation site. This intron contains several regulatory elements found in many eukaryotic genes. This sequence, when inserted upstream of a promoter-free luciferase gene, induces the expression of luciferase, demonstrating the promoter activity of the intron upon transfection in human or murine cells. This promoter activity is down-modulated by wild-type p53 but not by a mutated form of p53.

Antigens, Differentiation↗

An EBV-based vector allowing a high level of LTRHIV-directed expression in human cells.

We have developed a vector that allows high and transactivable expression of inserted genes. The vector contains a transcription unit in which the LTR from HIV flanks a multicloning site. The plasmid is based on the EBV p205 plasmid, which allows stable replication in human cells. The ability of the vector to express an exogenous DNA in human cells has been tested using the firefly luciferase gene.

DNA, Complementary↗

Specific gene transfer mediated by lactosylated poly-L-lysine into hepatoma cells.

Plasmid DNA/glycosylated polylysine complexes were used to transfer in vitro a luciferase reporter gene into human hepatoma cells by a receptor-mediated endocytosis process. HepG2 cells which express a galactose specific membrane lectin were efficiently and selectively transfected with pSV2Luc/lactosylated polylysine complexes in a sugar dependent manner: i) HepG2 cells which do not express membrane lectin specific for mannose were quite poorly transfected with pSV2Luc/mannosylated polylysine complexes, ii) HeLa cells which do not express membrane lectin specific for galactose were not transfected with pSV2Luc/lactosylated polylysine complexes. The transfection efficiency of HepG2 cells with pSV2Luc/lactosylated polylysine complexes was greatly enhanced either in the presence of chloroquine or in the presence of a fusogenic peptide. A 22-residue peptide derived from the influenza virus hemagglutinin HA2 N-terminal polypeptide that mimics the fusogenic activity of the virus, was selected. In the presence of the fusogenic peptide, the luciferase activity in HepG2 cells was 10 fold larger than that of cells transfected with pSV2Luc/lactosylated polylysine complexes in the presence of chloroquine.

Amino Acid Sequence↗

[Regulation of the biosynthesis of branched aminoacyl tRNA synthetases in Bacillus cereus T].

The regulation of the biosynthesis of isoleucyl-, valyl-, and leucyl-tRNA synthetases was studied with an isoleucine-valine requiring mutant of Bacillus cereus T. It was shown that valyl-tRNA synthetase regulation is under multivalent control involving both valine and isoleucine. The isoleucyl- and leucyl-tRNA synthetases are repressed by the respective cognate amino acid. When two amino acids were removed from the culture medium, derepression of the two corresponding aminoacyl tRNA synthetases was expected, but only one appears. With a threonine deaminase constitutive mutant, it was demonstrated that the derepression mecanism of the synthetase was correlated with the intracellular level of the ilv A gene product. These results are in good agreement with the model proposed by several authors. In this model, threonine deaminase, or some form of this enzyme, is involved in a positive control of the regulation of the branched-chain aminoacyltransfer ribonucleic acid synthetases.

Amino Acyl-tRNA Synthetases↗