title: Genomic organization and alternative transcripts of the human Connexin40 gene.
abstract: The human Cx40 gene (NT_004434.5) was sorted out from the GenBank database and as a result of a BLAST homology search, two ESTs (BE784549 from a human lung database, and BE732411 from a human placenta database) overlapping with the coding exon 2 sequence and upstream regions of the gene were identified. These ESTs correspond to two transcripts 1A and 1B, which diverge from each other in their 5' regions. The transcript 1A corresponds to the only transcript previously identified for the mouse and rat Cx40 genes; whereas the transcript 1B is a new transcript. The human Cx40 gene therefore comprises three exons: exon 1A (100 bp), exon 1B (132 bp) and coding exon 2, with the exons 1A and 1B at 14 and 1.3 kb of the exon 2, respectively. The expression of these transcripts is cell-type specific. Transcript 1A is expressed in endothelial cells. Its expression was demonstrated in human umbilical vein endothelial cells (HUVEC). Transcript 1B is expressed in placental cytotrophoblasts. Its expression was demonstrated in malignant trophoblastic cells, BeWo, JAR and JEG-3, and purified cytotrophoblasts from human first trimester placental tissues. Interestingly, both transcripts 1A and 1B are expressed in the right atrial appendages (RAA), although the cell-type expression of the two transcripts in this particular tissue has not yet been determined. Both transcripts were found to be expressed in the various heart regions investigated, where transcript 1B was found to always occur rarely in comparison with transcript 1A. Transcripts 1A and 1B are both more abundant in the atria than in the ventricles. Luciferase reporter gene assays demonstrated that two genomic regions containing the exons 1A and 1B induced a cell-type specific expression. The 1.2 kb sequence, containing the exon 1A, induced an increase of the luciferase activity in HUVEC; whereas the 1.9 kb sequence, containing the exon 1B, induces an increase of expression of the luciferase activity in BeWo cells. The DNA sequence upstream of the exon 1A contains SP1 binding sites, but no TATA- or CAAT-box; whereas the region upstream of the exon 1B is preceded by three CAAT-boxes. Thus, in contrast to the mouse and rat Cx40 genes, the human Cx40 gene organized in three exons and generates two transcripts, which are cell-type specific.
authors.affiliation: Laboratoire de Génétique et Physiologie du Développement, UMR 6545, Institut de Biologie du Développement de Marseille, Université de la Méditerranée, 13288 Marseille Cedex 9, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Dupays
authors.forename: David
authors.initials: D
authors.lastname: Mazurais
authors.forename: Catherine
authors.initials: C
authors.lastname: Rücker-Martin
authors.forename: Thierry
authors.initials: T
authors.lastname: Calmels
authors.forename: Denis
authors.initials: D
authors.lastname: Bernot
authors.forename: Laurent
authors.initials: L
authors.lastname: Cronier
authors.forename: André
authors.initials: A
authors.lastname: Malassiné
authors.forename: Daniel
authors.initials: D
authors.lastname: Gros
authors.forename: Magali
authors.initials: M
authors.lastname: Théveniau-Ruissy
mesh_terms: D024506:5' Flanking Region / Q000235:genetics; D020121:5' Untranslated Regions / Q000235:genetics / Q000378:metabolism; D017398:Alternative Splicing; D001483:Base Sequence; D002478:Cells, Cultured; D017630:Connexins / Q000235:genetics*; D004247:DNA / Q000737:chemistry / Q000235:genetics; D020224:Expressed Sequence Tags; D005260:Female; D005796:Genes / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011247:Pregnancy; D011401:Promoter Regions, Genetic / Q000235:genetics; D020033:Protein Isoforms / Q000235:genetics; D012313:RNA / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D024363:Transcription Initiation Site; D014158:Transcription, Genetic / Q000235:genetics; D014407:Tumor Cells, Cultured; D000097003:Gap Junction alpha-5 Protein
chemical_list: D020121:5' Untranslated Regions; D017630:Connexins; D004247:DNA; D008156:Luciferases; D020033:Protein Isoforms; D012313:RNA; D011993:Recombinant Fusion Proteins; D000097003:Gap Junction alpha-5 Protein