title: Cloning of novel LERGU mRNAs in GPR30 3' untranslated region and detection of 2 bp-deletion polymorphism in gastric cancer.
abstract: The improved IGCR (In-Gel Competitive Reassociation) method was applied to the analysis of human gastric cancer genomic DNA to identify its alterations, and it appeared that the IGCR library contained a fragment of 3'-untranslated region (3' UTR) of G-protein coupled receptor 30 (GPR30) mRNA. When we searched genomic DNA pairs of gastric cancer patients with this IGCR clone, we found the deletion polymorphism with or without 2 bp (Cytosine and Thymine; CT). We confirmed the existence of a novel mRNA in GPR30 3'UTR by northern blotting, cloned this novel mRNA and named it Leucine Rich Protein in GPR30 3'UTR (LERGU). The EST database search gave one alternative splicing form in this 3' UTR, which was named as LERGU-1. A novel alternative splicing form of this mRNA was also identified from the stomach total RNA, which was named LERGU-2. The LERGU mRNA was also detected in eight gastric cancer cell lines, but GPR30 mRNA scarcely existed. Furthermore, we detected the 2 bp-deletion form in genomic DNAs and mRNAs derived from gastric cancers, but not in other type cancers. Since the 2 bp-deletion position on LERGU corresponds to its alternative splicing site, this deletion may produce a frame-shifted protein. Overall, our findings suggest that a mutation or disappearance of the normal LERGU protein may have a function in the development of gastric cancer.
authors.affiliation: Tsukuba Research Institute, Novartis Pharma K.K., Ohkubo 8, Tsukuba-shi Ibaraki 300-2611, Japan.
authors.forename: Naoto
authors.initials: N
authors.lastname: Kakinuma
authors.forename: Masaaki
authors.initials: M
authors.lastname: Sato
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Yamada
authors.forename: Kazuyoshi
authors.initials: K
authors.lastname: Kohu
authors.forename: Motowo
authors.initials: M
authors.lastname: Nakajima
authors.forename: Tetsu
authors.initials: T
authors.lastname: Akiyama
authors.forename: Susumu
authors.initials: S
authors.lastname: Ohwada
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Shibanaka
mesh_terms: D020413:3' Untranslated Regions; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics*; D016368:Frameshift Mutation; D006801:Humans; D007930:Leucine; D008969:Molecular Sequence Data; D011110:Polymorphism, Genetic; D012333:RNA, Messenger / Q000032:analysis; D011960:Receptors, Estrogen; D043562:Receptors, G-Protein-Coupled / Q000235:genetics*; D013274:Stomach Neoplasms / Q000235:genetics* / Q000473:pathology
chemical_list: D020413:3' Untranslated Regions; D004273:DNA, Neoplasm; C109584:GPER1 protein, human; D012333:RNA, Messenger; D011960:Receptors, Estrogen; D043562:Receptors, G-Protein-Coupled; D007930:Leucine