Novel Polymorphisms and Genetic Characteristics of the Prion Protein Gene (PRNP) in Dogs—A Resistant Animal of Prion Disease

Dong-Ju Kim,An-Dang Kim,Byung-Hoon Jeong,Yong-Chan Kim

doi:10.3390/ijms21114160

Abstract

Transmissible spongiform encephalopathies (TSEs) have been reported in a wide range of species. However, TSE infection in natural cases has never been reported in dogs. Previous studies have reported that polymorphisms of the prion protein gene (PRNP) have a direct impact on the susceptibility of TSE. However, studies on polymorphisms of the canine PRNP gene are very rare in dogs. We examined the genotype, allele, and haplotype frequencies of canine PRNP in 204 dogs using direct sequencing and analyzed linkage disequilibrium (LD) using Haploview version 4.2. In addition, to evaluate the impact of nonsynonymous polymorphisms on the function of prion protein (PrP), we carried out in silico analysis using PolyPhen-2, PROVEAN, and PANTHER. Furthermore, we analyzed the structure of PrP and hydrogen bonds according to alleles of nonsynonymous single nucleotide polymorphisms (SNPs) using the Swiss-Pdb Viewer program. Finally, we predicted the impact of the polymorphisms on the aggregation propensity of dog PrP using AMYCO. We identified a total of eight polymorphisms, including five novel SNPs and one insertion/deletion polymorphism, and found strong LDs and six major haplotypes among eight polymorphisms. In addition, we identified significantly different distribution of haplotypes among eight dog breeds, however, the kinds of identified polymorphisms were different among each dog breed. We predicted that p.64_71del HGGGWGQP, Asp182Gly, and Asp182Glu polymorphisms can impact the function and/or structure of dog PrP. Furthermore, the number of hydrogen bonds of dog PrP with the Glu182 and Gly182 alleles were predicted to be less than those with the Asp182 allele. Finally, Asp163Glu and Asp182Gly showed more aggregation propensity than wild-type dog PrP. These results suggest that nonsynonymous SNPs, Asp182Glu and Asp182Gly, can influence the stability of dog PrP and confer the possibility of TSE infection in dogs.

Highlights

Transmissible spongiform encephalopathies (TSEs), known as prion diseases, are neurodegenerative diseases caused by conversion of the normal prion protein (PrPC) into aggregated, self-propagating and disease-associated isoforms (PrPSc)
We evaluated the impact of polymorphisms of the canine prion protein gene (PRNP) gene on dog PrP using PolyPhen-2, PROVEAN, PANTHER, and AMYCO [33,34,35,36,37]
We compared the distribution of most haplotype 1 of the canine PRNP gene among eight dog breeds

Summary

Introduction

Transmissible spongiform encephalopathies (TSEs), known as prion diseases, are neurodegenerative diseases caused by conversion of the normal prion protein (PrPC) into aggregated, self-propagating and disease-associated isoforms (PrPSc). Dogs were likely to have been exposed to BSE-contaminated food, TSE infection was never reported in dogs [12,13]. A circular dichroism study reported that the PrP of dogs is more stable than that of prion disease-susceptible animals, including hamsters and mice, at four pH values [16]. In a study using protein misfolding cyclic amplification (PMCA), dog brain homogenate showed resistance to several prion agents, including BSE, scrapie, and chronic wasting disease (CWD) [19]. The dog-specific amino acid (Asp163), which is located on the α1-β2 loop of PrP-transgenic mice, showed complete resistance to TSEs following intracerebral inoculation with three mouse-adapted scrapie strains, including Rocky Mountain Laboratory (RML), 301C, and 22L [18]

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Results

Conclusion