title: hSmad5 gene, a human hSmad family member: its full length cDNA, genomic structure, promoter region and mutation analysis in human tumors.
abstract: hSmad (mothers against decapentaplegic)-related proteins are important messengers within the Transforming Growth Factor-beta1 (TGF-beta1) superfamily signal transduction pathways. To further characterize a member of this family, we obtained a full length cDNA of the human hSmad5 (hSmad5) gene by rapid amplification of cDNA ends (RACE) and then determined the genomic structure of the gene. There are eight exons and two alternative transcripts; the shorter transcript lacks exon 2. We identified the hSmad5 promoter region from a human genomic YAC clone by obtaining the nucleotide sequence extending 1235 base pairs upstream of the 5' end of the cDNA. We found a CpG island consistent with a promoter region, and we demonstrated promoter activity in a 1232 bp fragment located upstream of the transcription initiation site. To investigate the frequency of somatic hSmad5 mutations in human cancers, we designed intron-based primers to examine coding regions by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis. Neither homozygous deletions or point mutations were found in 40 primary gastric tumors and 51 cell lines derived from diverse types of human cancer including 20 cell lines resistant to the growth inhibitory effects of TGF-beta1. These results suggest that the hSmad5 gene is not commonly mutated and that other genetic alterations mediate the loss of TGF-beta1 responsiveness in human cancers.
authors.affiliation: Laboratory of Human Carcinogenesis, Division of Basic Sciences, National Cancer Institute, NIH, Bethesda, Maryland 20892-4255, USA.
authors.forename: A
authors.initials: A
authors.lastname: Gemma
authors.forename: K
authors.initials: K
authors.lastname: Hagiwara
authors.forename: F
authors.initials: F
authors.lastname: Vincent
authors.forename: Y
authors.initials: Y
authors.lastname: Ke
authors.forename: A R
authors.initials: AR
authors.lastname: Hancock
authors.forename: M
authors.initials: M
authors.lastname: Nagashima
authors.forename: W P
authors.initials: WP
authors.lastname: Bennett
authors.forename: C C
authors.initials: CC
authors.lastname: Harris
mesh_terms: D017398:Alternative Splicing; D000595:Amino Acid Sequence; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D004273:DNA, Neoplasm / Q000235:genetics; D004268:DNA-Binding Proteins; D005796:Genes; D006801:Humans; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D016415:Sequence Alignment; D051902:Smad5 Protein; D015534:Trans-Activators; D014407:Tumor Cells, Cultured
chemical_list: D018076:DNA, Complementary; D004273:DNA, Neoplasm; D004268:DNA-Binding Proteins; D010750:Phosphoproteins; D012333:RNA, Messenger; C497797:SMAD5 protein, human; D051902:Smad5 Protein; D015534:Trans-Activators