NR AYGA
AU Nakamura,I.; Xue,G.; Sakudo,A.; Saeki,K.; Matsumoto,Y.; Ikuta,K.; Onodera,T.
TI Novel single nucleotide polymorphisms in the specific protein 1 binding site of the bovine PRNP promoter in Japanese Black cattle: impairment of its promoter activity.
QU Intervirology 2007; 50(3): 190-6
PT comparative study; journal article; research support, non-u.s. gov't
AB Susceptibility to transmissible spongiform encephalopathy and different alleles of the prion protein gene (PRNP) of humans and sheep are associated. A tentative association between PRNP promoter polymorphisms and bovine spongiform encephalopathy (BSE) susceptibility has been reported in German cattle, whereas none of the known polymorphisms within the bovine PRNP-coding sequence affect BSE susceptibility. In the present study, novel single nucleotide polymorphisms located in the 5'-flanking region of bovine PRNP affecting its expression were demonstrated in Japanese Black cattle. We sequenced exon 1, and the approximately 200-bp 5'-flanking region of the PRNP translation initiation site containing the proximal promoter of PRNP was harvested. We identified 7 single nucleotide polymorphisms: -184A-->G, -141T-->C, -85T-->G, -47C-->A, -6C-->T, +17C-->T and +43C-->T. Six segregated haplotypes in the population were cloned into luciferase-expressing plasmids, transfected into N2a cells, and their reporter activities were measured 48 h after transfection. Six haplotypes showed a decreased expression level including -6C-->T in specific protein 1 binding site (p < 0.05) or -141T-->C (p < 0.01) at 48 h compared with the wild-type haplotype. These results advocate that certain polymorphisms such as specific protein 1 binding site polymorphisms in the bovine PRNP promoter region in Japanese Black cattle could influence promoter activity, suggesting that breeding cattle with such substitutions may be a useful approach in reducing BSE risk.
MH 5' Flanking Region/genetics; Animals; Base Sequence; Binding Sites/genetics; Cattle; Cloning, Molecular; Encephalopathy, Bovine Spongiform/*genetics; Gene Expression; Genes, Reporter; *Genetic Predisposition to Disease; Haplotypes; Luciferases/biosynthesis; Molecular Sequence Data; *Polymorphism, Single Nucleotide; Prions/*genetics; *Promoter Regions (Genetics); Sequence Analysis, DNA
AD Department of Molecular Immunology, School of Agricultural and Life Sciences, University of Tokyo, Tokyo, Japan.
SP englisch
PO Schweiz