title: Characterization of the human porin isoform 1 (HVDAC1) gene by amplification on the whole human genome: A tool for porin deficiency analysis.
abstract: The deficiency of porin isoform 1 (HVDAC1) in human skeletal muscle has been associated with a pathological phenotype related to defects in the bioenergetic metabolism. In the best studied case, porin deficiency was not apparent in cultured fibroblasts: this observation raised the conclusion that no molecular defect was in the cDNA sequence coding for the protein. To get more insight in the pathogenetic mechanism that is involved in porin isoform 1 deficiency, we have determined the whole structure of the corresponding human gene. On the basis of the corresponding mouse gene structure and the human cDNA sequence, we designed long extension PCR amplifications using the whole genomic DNA as a template. Exonic/intronic regions were isolated and the exons and surrounding introns sequenced. The 5' and 3' extremities of the gene were determined by genome walking. The porin isoform 1 human gene is made up of 9 exons and spans about 33 kbp. A whole panel of PCR parameters was set and is now ready to be used for specific amplification upon patients' genomic DNA. The analysis of the putative promoter sequence was performed. It revealed the presence of a sterol Repressor element (SRE), an SRY, the testis-determining factor, and a nuclear respiratory factor 2 (NRF-2) binding site. These sites, according to results from literature, could be involved in the functional modulation of the gene expression.
authors.affiliation: Department of Chemical Sciences, Laboratory of Biochemistry and Molecular Biology, Università di Catania, viale A. Doria 6, Catania, I-95125, Italy. mess@mbox.unict.it
authors.forename: A
authors.initials: A
authors.lastname: Messina
authors.forename: F
authors.initials: F
authors.lastname: Guarino
authors.forename: M
authors.initials: M
authors.lastname: Oliva
authors.forename: L P
authors.initials: LP
authors.lastname: van den Heuvel
authors.forename: J
authors.initials: J
authors.lastname: Smeitink
authors.forename: V
authors.initials: V
authors.lastname: De Pinto
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D020121:5' Untranslated Regions / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D016386:Chromosome Walking; D003001:Cloning, Molecular; D005091:Exons; D005347:Fibroblasts / Q000378:metabolism; D005784:Gene Amplification; D015894:Genome, Human; D006801:Humans; D007438:Introns; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D018272:Porins / Q000172:deficiency* / Q000235:genetics*; D011401:Promoter Regions, Genetic; D020033:Protein Isoforms / Q000235:genetics; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D050995:Voltage-Dependent Anion Channel 1; D050994:Voltage-Dependent Anion Channels
chemical_list: D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D018272:Porins; D020033:Protein Isoforms; C494762:VDAC1 protein, human; D050994:Voltage-Dependent Anion Channels; D050995:Voltage-Dependent Anion Channel 1