title: MiR-128 and miR-125 regulate expression of coagulation Factor IX gene with nonsense mutation by repressing nonsense-mediated mRNA decay.
abstract: Hemophilia could be caused by a nonsense mutation of the Factor IX gene, leading to a deficiency of Factor IX (F9). The nonsense mutation frequency of F9 is more than 10% according to the database. Nonsense-mediated mRNA decay (NMD) is a defined cellular response that can potentially prevent the production of such deleterious C-terminal truncated proteins from aberrant mRNA. Here, we constructed a mini-gene of Factor IX (Mini-hF9) and some nonsense mutants and characterized the mini-gene splicing pattern. We discovered that NMD regulated mini-hF9 expression in two nonsense mutants: E7a (nt 34 G>T in exon 7) and E7b (nt 52 G>T in exon 7), but not in another nonsense mutants: E7c (nt 85 G>T in exon 7) and E8 (nt 42 C>T in exon 8). In addition, mini-hF9 transcripts were accumulated after transfection with miR-128 or miR-125 mimics in E7a and E7b. Our results suggest that PTC (premature termination codon) location is a key determination for triggering NMD; miR-128 and miR-125 could help to increase the nonsense-mutant F9 levels by repressing NMD.
authors.affiliation: Department of Hematology, The Second Clinical Medical College, Shanxi Medical University, Taiyuan 030001, PR China.
authors.forename: Gang
authors.initials: G
authors.lastname: Wang
authors.affiliation: Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan 030006, PR China. Electronic address: bfchai@sxu.edu.cn.
authors.forename: Baofeng
authors.initials: B
authors.lastname: Chai
authors.affiliation: Department of Hematology, The Second Clinical Medical College, Shanxi Medical University, Taiyuan 030001, PR China. Electronic address: yanglh5282@163.com.
authors.forename: Linhua
authors.initials: L
authors.lastname: Yang
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D018389:Codon, Nonsense / Q000235:genetics*; D005164:Factor IX / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D059365:Nonsense Mediated mRNA Decay / Q000235:genetics*
chemical_list: D018389:Codon, Nonsense; C531888:MIRN125 microRNA, human; C545189:MIRN128 microRNA, human; D035683:MicroRNAs; D005164:Factor IX
keywords: Factor IX; Hemophilia; MicroRNA; NMD; Nonsense mutation