NR ATMN
AU Kashkevich,K.; Humeny,A.; Sander,P.; Orlicz-Welcz,B.; Henz,I.; Leeb,T.; Becker,C.M.; Schiebel,K.
TI Polymorphisms of the prion protein gene promoter region and their association with BSE
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Genetics, strains and emerging problems GEN-13
PT Konferenz-Poster
AB
DNA polymorphisms of the prion protein gene (PRNP) in humans and sheep are known to be associated with varying degrees of susceptibility to transmissible spongiform encephalopathies. However, no polymorphism within the bovine PRNP coding region has been shown to be associated with bovine spongiform encephalopathy (BSE). To determine whether polymorphisms within the promoter region are involved in susceptibility or resistance to BSE, a three-step approach was used. Initially, DNA-sequencing of the promoter region of the prion protein gene from control and BSE-affected animals was performed to assess the degree of genetic variation between different bovine breeds. As a second step, the functionality of a 12 bp indel was assessed using a luciferase reporter gene assay. Finally, the binding of different transcriptional factors to DNA fragments in which polymorphisms were detected was examined.
Comparative sequencing between control and BSE-affected animals from 4 different bovine breeds (namely Schwarzbunt, Rotbunt, Braunvieh and Fleckvieh) showed that Braunvieh animals are significantly different from other breeds, exhibiting different allele frequencies for the 12 bp indel polymorphism both in control and BSE animals. The deletion or insertion of the 12 bp fragment respectively deletes or creates a potential Sp1 binding site. Reporter gene assay experiments comparing both alleles resulted in different expression of constructs. Furthermore, using an electrophoretic mobility shift assay different transcription factor binding of the insertion and deletion alleles could be demonstrated. Binding of both, Sp1 and Sp3 transcription factors to the ins-allele indicated that expression of the ins-allele not only depends on the activation of Sp1 but also on the proportion of Sp1 to Sp3 since Sp3 may act as a repressor of transcription.
AD K.Kashkevich, A.Humeny, B.Orlicz-Welcz, I.Henz, C.-M.Becker, K.Schiebel, Institut für Biochemie, Universität Erlangen-Nürnberg, Germany; P.Sander, T.Leeb, Institut füt Tierzucht und Genetik, Tiermedizinische Hochschule Hannover, Germany
SP englisch
PO Deutschland