title: Brain- and heart-specific Patched-1 containing exon 12b is a dominant negative isoform and is expressed in medulloblastomas.
abstract: Mutations in the human tumor suppressor gene, Patched-1, are associated with nevoid basal cell carcinoma syndrome characterized by developmental abnormalities and tumorigenesis, such as basal cell carcinoma and medulloblastoma. During the investigation of complex alternative splicing in Patched-1, we identified an alternative exon, exon 12b, located between exon 12 and 13, both in humans and in mice. Since exon 12b has an in-frame stop codon, the mRNA isoform containing this exon (Patched12b) encodes a truncated patched-1 protein. RT-PCR and whole mount in situ hybridization revealed that mouse exon 12b was expressed in the brain and heart, particularly in the cerebellum, in both adults and embryos. We next performed a functional analysis of Patched12b using a GLI-responsive luciferase reporter. Luciferase activity was suppressed when transfected with a plasmid encoding Patched-1, but not with a plasmid for Patched12b. The suppressive activity of Patched-1 was relieved when cotransfected with a plasmid for Patched12b. This implies that the Patched12b protein has a dominant negative effect on Patched-1. Interestingly, Patched12b was found to be expressed in some of the medulloblastoma tissues and cell lines, indicating an important role in the pathogenesis of medulloblastoma as well as brain development.
authors.affiliation: Department of Genetics, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan.
authors.forename: Hideki
authors.initials: H
authors.lastname: Uchikawa
authors.forename: Masashi
authors.initials: M
authors.lastname: Toyoda
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Nagao
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Miyauchi
authors.forename: Ryo
authors.initials: R
authors.lastname: Nishikawa
authors.forename: Katsunori
authors.initials: K
authors.lastname: Fujii
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Kohno
authors.forename: Masao
authors.initials: M
authors.lastname: Yamada
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Miyashita
mesh_terms: D017398:Alternative Splicing; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D005091:Exons; D015972:Gene Expression Regulation, Neoplastic; D005799:Genes, Dominant; D006801:Humans; D008527:Medulloblastoma / Q000378:metabolism*; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism*; D000072079:Patched Receptors; D000072081:Patched-1 Receptor; D020033:Protein Isoforms; D011956:Receptors, Cell Surface / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D014018:Tissue Distribution
chemical_list: C000606236:PTCH1 protein, human; D000072079:Patched Receptors; D000072081:Patched-1 Receptor; D020033:Protein Isoforms; C000606240:Ptch1 protein, mouse; D011956:Receptors, Cell Surface