Messenger RNA transcripts of the hepatocyte nuclear factor-1alpha gene containing premature termination codons are subject to nonsense-mediated decay.
Mutations in the hepatocyte nuclear factor-1α (HNF1a) gene cause maturity-onset diabetes of the young (MODY). Approximately 30% of these mutations generate mRNA transcripts harboring premature termination codons (PTCs). Degradation of such transcripts by the nonsense-mediated decay (NMD) pathway has...
| Publicado en: | Diabetes Vol. 53; no. 2; pp. 500 - 505 |
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| Autores principales: | , , |
| Formato: | research Journal Article |
| Publicado: |
American Diabetes Association
Feb2004
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=12209457&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 12209457 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 00121797 1G6 jtl: Diabetes issn: 00121797 maglogo: N pubinfo: dt: Feb2004 vid: 53 iid: 2 pid: 1367 pub: American Diabetes Association place: Arlington, Virginia artinfo: ui: 12209457 12209457 NLM14747304 12209457 10.2337/diabetes.53.2.500 NLM14747304 12209457 ppf: 500 ppct: 5 formats: tig: atl: Messenger RNA transcripts of the hepatocyte nuclear factor-1alpha gene containing premature termination codons are subject to nonsense-mediated decay. aug: au: Harries, Lorna W. Hattersley, Andrew T. Ellard, Sian affil: From the Institute of Biomedical and Clinical Science, Peninsula Medical School, Exeter, U.K. sug: subj: RNA Mutation DNA-Binding Proteins Diabetes Mellitus, Type 2 Nuclear Proteins Transcription Factors RNA Drug Effects DNA Probes Genes Nucleotides Amino Acids Family Proteins Reference Values Human Reverse Transcriptase Polymerase Chain Reaction Cell Line Disease Susceptibility Piperidines Pharmacodynamics Validation Studies Comparative Studies Evaluation Research Multicenter Studies ab: Mutations in the hepatocyte nuclear factor-1α (HNF1a) gene cause maturity-onset diabetes of the young (MODY). Approximately 30% of these mutations generate mRNA transcripts harboring premature termination codons (PTCs). Degradation of such transcripts by the nonsense-mediated decay (NMD) pathway has been reported for many genes. To determine whether PTC mutant transcripts of the HNF-1α gene elicit NMD, we have developed a novel quantitative RT-PCR assay. We performed quantification of ectopically expressed murant transcripts relative to normal transcripts in lymphoblastoid cell lines using a coding single nucleotide polymorphism (cSNP) as a marker. The nonsense mutations R171X, I414G415ATCG→CCA, and P291fsinsC showed reduced mutant mRNA expression to 40% (P = 0.009), <0.01% (P ≤ 0.0001), and 6% (P = 0.001), respectively, of the normal allele. Transcript levels were restored using the translation inhibitor cycloheximide, indicating that the instability arises from NMD. The missense mutations G207D and R229P did not show NMD although R229P exhibited moderate RNA instability. This study provides the first evidence that HNF-1α PTC mutations may be subject to NMD. Mutations that result in significant reduction of protein levels due to NMD will not have dominant-negative activity in vivo. Haploinsufficiency is therefore likely to be the most important mutational mechanism of HNF-1α mutations causing MODY. pubtype: Academic Journal doctype: research Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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