Cofactor and glycosylation preferences for in vitro prion conversion are predominantly determined by strain conformation
| Autor: | Kenneth M. K. Mark, Nathan R. Deleault, Umberto Agrimi, Koren Nishina, Daniel J. Walsh, Surachai Supattapone, Michele Angelo Di Bari, Cassandra M. Burke |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Glycosylation
PrPSc Proteins animal diseases Molecular Conformation Glycobiology Protein Sequencing Biochemistry Prion Diseases Mice chemistry.chemical_compound Protein sequencing Cricetinae Zoonoses Medicine and Health Sciences Post-Translational Modification Biology (General) Enzyme Chemistry Peptide sequence Phospholipids Mammals chemistry.chemical_classification 0303 health sciences biology Strain (chemistry) Arvicolinae Biochemical Cofactors 030302 biochemistry & molecular biology Brain Eukaryota Animal Models Lipids Infectious Diseases Experimental Organism Systems Vertebrates Hamsters Research Article Prions QH301-705.5 Immunology Mouse Models Research and Analysis Methods Communicable Diseases Rodents Microbiology Cofactor 03 medical and health sciences Model Organisms Species Specificity Virology mental disorders Genetics Animals PrPC Proteins Amino Acid Sequence Molecular Biology Techniques Sequencing Techniques Molecular Biology 030304 developmental biology Mesocricetus Voles Organisms Biology and Life Sciences Proteins RNA RC581-607 In vitro nervous system diseases Mice Inbred C57BL chemistry Amniotes Enzymology Animal Studies biology.protein Parasitology Immunologic diseases. Allergy Glycoprotein |
| Zdroj: | PLoS Pathogens, Vol 16, Iss 4, p e1008495 (2020) PLoS Pathogens |
| ISSN: | 1553-7374 1553-7366 |
| Popis: | Prion diseases are caused by the misfolding of a host-encoded glycoprotein, PrPC, into a pathogenic conformer, PrPSc. Infectious prions can exist as different strains, composed of unique conformations of PrPSc that generate strain-specific biological traits, including distinctive patterns of PrPSc accumulation throughout the brain. Prion strains from different animal species display different cofactor and PrPC glycoform preferences to propagate efficiently in vitro, but it is unknown whether these molecular preferences are specified by the amino acid sequence of PrPC substrate or by the conformation of PrPSc seed. To distinguish between these two possibilities, we used bank vole PrPC to propagate both hamster or mouse prions (which have distinct cofactor and glycosylation preferences) with a single, common substrate. We performed reconstituted sPMCA reactions using either (1) phospholipid or RNA cofactor molecules, or (2) di- or un-glycosylated bank vole PrPC substrate. We found that prion strains from either species are capable of propagating efficiently using bank vole PrPC substrates when reactions contained the same PrPC glycoform or cofactor molecule preferred by the PrPSc seed in its host species. Thus, we conclude that it is the conformation of the input PrPSc seed, not the amino acid sequence of the PrPC substrate, that primarily determines species-specific cofactor and glycosylation preferences. These results support the hypothesis that strain-specific patterns of prion neurotropism are generated by selection of differentially distributed cofactors molecules and/or PrPC glycoforms during prion replication. Author summary According to the “protein-only hypothesis,” mammalian prions are unconventional infectious agents that lack replicating nucleic acids and instead contain misfolded forms of a host glycoprotein termed PrPSc. Paradoxically, despite lacking independent genomes, prions can exist as distinct self-propagating “strains,” each of which is associated with its own PrPSc conformation and biological properties, including unique patterns of brain targeting (neurotropism) and PrPSc glycosylation. The mechanism by which different PrPSc conformers can cause distinct patterns of neurotropism and PrPSc glycosylation is unknown, and represents an important challenge for the protein-only hypothesis. Here, we show that the prion strain conformation plays a dominant role in determining which cofactor molecules and glycosylated substrate molecules can be used to form PrPSc in chemically defined biochemical assays. These results provide the first direct evidence that the major strain properties of infectious prions, including neurotropism, can be explained by the process of selective cofactor and substrate usage during PrPSc replication. This concept may also explain the specific patterns of neurotropism observed for several other prion-like neurodegenerative diseases, such as Parkinson’s disease and Alzheimer’s disease. |
| Databáze: | OpenAIRE |
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