کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5905697 | 1159926 | 2014 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Defective pre-mRNA splicing in PKD1 due to presumed missense and synonymous mutations causing autosomal dominant polycystic disease
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کلمات کلیدی
μLESEADPKDdNTPGenetic kidney diseaseNMDISSESSExonic mutationRT-PCRDMEMcDNA - cDNADNA complementary to RNA - DNA مکمل RNAISE - ISADulbecco's modified Eagle's medium - Medal of Eagle اصلاح شده Dulbeccoautosomal dominant polycystic kidney disease - بیماری کلیوی پلی کیستیک غالب در اتواسوم غالب استExonic splicing enhancers - تقویت کننده های اسپلایسینگ اکسونbase pairs - جفت پایهMin - حداقلdeoxyribonucleoside triphosphate - دز اکسید ریبونوکلئوزید تری فسفاتminutes - دقایقDonor splice site - سایت اسپلایس دونرExonic splicing silencers - صدا خفه کن های اسپلیکسونnonsense-mediated mRNA decay - فرسودن mRNA ناشی از بی معنی استLuria–Bertani (medium) - لوریا بارتانی (متوسط)millimolar - میلی آمپرMicroliter - میکرولیترwild-type - نوع وحشیUnits - واحدهاpolymerase chain reaction - واکنش زنجیره ای پلیمرازreverse transcription polymerase chain reaction - واکنش زنجیره ای پلیمراز رونویسی معکوسPCR - واکنش زنجیرهٔ پلیمراز
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
ژنتیک
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Defective pre-mRNA splicing in PKD1 due to presumed missense and synonymous mutations causing autosomal dominant polycystic disease Defective pre-mRNA splicing in PKD1 due to presumed missense and synonymous mutations causing autosomal dominant polycystic disease](/preview/png/5905697.png)
چکیده انگلیسی
Autosomal dominant polycystic kidney disease is the most common human monogenic disorder and is caused by mutations in the PKD1 or PKD2 genes. Most patients with the disease present mutations in PKD1, and a considerable number of these alterations are single base substitutions within the coding sequence that are usually predicted to lead to missense or synonymous mutations. There is growing evidence that some of these mutations can be detrimental by affecting the pre-mRNA splicing process. The aim of our study was to test PKD1 mutations, described as missense or synonymous in the literature or databases, for their effects on exon inclusion. Bioinformatics tools were used to select mutations with a potential effect on pre-mRNA splicing. Mutations were experimentally tested using minigene assays. Exons and adjacent intronic sequences were PCR-amplified and cloned in the splicing reporter minigene, and selected mutations were introduced by site-directed mutagenesis. Minigenes were transfected into kidney derived cell lines. RNA from cultured cells was analyzed by RT-PCR and DNA sequencing. Analysis of thirty-three PKD1 exonic mutations revealed three mutations that induce splicing defects. The substitution c.11156GÂ >Â A, previously predicted as missense mutation p.R3719Q, abolished the donor splice site of intron 38 and resulted in the incorporation of exon 38 with 117Â bp of intron 38 and skipping of exon 39. Two synonymous variants, c.327AÂ >Â T (p.G109G) and c.11257CÂ >Â A (p.R3753R), generated strong donor splice sites within exons 3 and 39 respectively, resulting in incorporation of incomplete exons. These three nucleotide substitutions represent the first PKD1 exonic mutations that induce aberrant mRNAs. Our results strengthen the importance to evaluate the consequences of presumed missense and synonymous mutations at the mRNA level.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Gene - Volume 546, Issue 2, 10 August 2014, Pages 243-249
Journal: Gene - Volume 546, Issue 2, 10 August 2014, Pages 243-249
نویسندگان
Francisco J. Gonzalez-Paredes, Elena Ramos-Trujillo, Felix Claverie-Martin,