title: The porcine sodium/iodide symporter gene exhibits an uncommon expression pattern related to the use of alternative splice sites not present in the human or murine species.
abstract: The sodium/iodide symporter (NIS) is a membrane protein mediating the active transport of iodide into the thyroid gland. NIS, expressed by human, rat, and mouse thyrocytes, is encoded by a single transcript. We identified NIS mRNA species of 3.5 and 3 kb in porcine thyrocytes. Because porcine thyrocytes in primary culture is a widely used experimental system for thyroid iodide metabolism, we further examined the origin and the function of the porcine NIS (pNIS) transcripts. We generated a porcine thyroid cDNA library from which four different clones, pNIS-D, F, J, and Delta J were isolated. pNIS-D encodes a protein of 643 amino acids highly homologous to the human, rat, and mouse NIS. pNIS-F and J differ from each other and from pNIS-D in their C-terminal part. pNIS-Delta J lacks a six-amino-acid segment within the putative transmembrane domain 10. Transiently expressed in Cos-7 cells, the four pNIS-cDNAs led to the synthesis of proteins targeted at the plasma membrane and conferred perchlorate-sensitive iodide uptake activities to Cos-7 cells, except pNIS-Delta J, which was devoid of activity. PNIS-D probably derives from the 3.5-kb transcript and pNIS-F, J, and Delta J from the 3-kb transcript. The relative abundance of pNIS-D, F, and J transcripts in porcine thyrocytes was about 60%, 35%, and 5%, respectively; the Delta J transcript was not present in detectable amount. By comparing porcine NIS genomic and cDNA sequences, splice donor and acceptor sites accounting for the generation of pNIS-F, J, and Delta J transcripts were identified. None of the combinations of alternative splice sites found in the pig was present in the human, rat or mouse NIS gene. Our data show that porcine NIS gene, contrary to the NIS gene from other species, gives rise to splice variants leading to three active and one inactive NIS proteins.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, Unité 369, Institut Fédératif de Recherche Laennec, 69372 Lyon, Cedex 08, France.
authors.forename: Samia
authors.initials: S
authors.lastname: Selmi-Ruby
authors.forename: Chantal
authors.initials: C
authors.lastname: Watrin
authors.forename: Severine
authors.initials: S
authors.lastname: Trouttet-Masson
authors.forename: Françoise
authors.initials: F
authors.lastname: Bernier-Valentin
authors.forename: Virginie
authors.initials: V
authors.lastname: Flachon
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Munari-Silem
authors.forename: Bernard
authors.initials: B
authors.lastname: Rousset
mesh_terms: D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D019556:COS Cells; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis / Q000737:chemistry; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D015723:Gene Library; D006801:Humans; D007454:Iodides / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D016415:Sequence Alignment; D013045:Species Specificity; D013552:Swine / Q000235:genetics*; D027981:Symporters / Q000737:chemistry / Q000235:genetics*; D013961:Thyroid Gland / Q000737:chemistry; D014162:Transfection; D000099209:Sodium-Iodide Symporters
chemical_list: D018076:DNA, Complementary; D007454:Iodides; D020033:Protein Isoforms; D012333:RNA, Messenger; D027981:Symporters; D000099209:Sodium-Iodide Symporters