NR AWFP
AU Gilliland,S.; Skogtvedt,S.; Tranulis,M.A.; Kristensen,T.; Goldmann,W.
TI Determining the structure and sequence of ovine genes which encode potential PrP binding proteins
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions GEN-11
PT Konferenz-Poster
AB The PrP protein gene is a major controlling factor of scrapie. Polymorphisms in the PrP gene lead to modulation of disease phenotypes. The mechanisms by which these PrP polymorphisms achieve their effect are still unknown but it is likely that interactions with other host proteins are of importance. We have begun to analyse genes of proteins that have been shown by in vitro studies to bind PrP. We have selected genes such as 140kDa NCAM, 37kDa laminin receptor precursor (LRP) and Pint1 for a detailed analysis of their gene structure and genetics in sheep, a natural host of classical and atypical scrapie. Here we report our data on ovine Pint1. Spielhaupter & Schätzl, (J Biol Chem. 276:44604-44612; 2001) described a prion interactor (Pint1) gene based on a partial mouse brain cDNA expressing a peptide domain that bound to PrP protein in a yeast two-hybrid system. A homologous cDNA is also present in human brain. To be able to analyse this protein interaction in sheep, we have cloned and sequenced Pint1-coding cDNAs from ovine brain and testis. The exon-intron structure of sheep Pint1 was inferred from comparison with the published mouse and human Pint1 gene structures and confirmed in part on sheep genomic clones. Our data reveal novel Pint1 mRNA variants in brain and testis derived through alternative splicing. Some of these alternative mRNAs encode variants in the Pint1 coding region, in one case this change is close to the putative PrP binding domain of Pint1. The PrP-binding domain was sequenced for brain cDNA from sheep, goat, cattle and mouse to analyse species specificities in this protein domain. The expression of Pint1 mRNA from sheep brain was quantified and compared with mouse. We conclude that the roles of Pint1 in ruminants and rodents in scrapie may differ
AD S. Gilliland, W. Goldmann: Institute for Animal Health, Neuropathogenesis Unit, Edinburgh, UK; S. Skogtved, M.A. Tranulis: Department of Basic Sciences and Aquatic Medicine, Norwegian School of Veterinary Science, Oslo, Norway; T. Kristensen: Department of Molecular Biosciences, University of Oslo, P.O.Box 1041 Blindern, Oslo, Norway
SP englisch
PO Italien