Cryo-EM structure of anchorless RML prion reveals variations in shared motifs between distinct strains

Hoyt, Forrest; Standke, Heidi G.; Artikis, Efrosini; Schwartz, Cindi L.; Hansen, Bryan; Li, Kunpeng; Hughson, Andrew G.; Manca, Matteo; Thomas, Olivia R.; Raymond, Gregory J.; Race, Brent; Baron, Gerald S.; Caughey, Byron; Kraus, Allison

doi:10.1038/s41467-022-30458-6

Cited by 61 publications

(84 citation statements)

References 37 publications

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“…These results show that the a22L preparation was highly infectious. Given that incubation period correlates inversely with prion titer for a given prion strain in a given type of host (23), the similarity of these incubation periods suggests that the a22L preparation contains roughly 3 x 10 10 LD50/mg, which is comparable to that of our previously titered aRML preparation (5). However, a caveat to this a22L titer estimate is that the purified a22L preparation and wildtype 22L in brain homogenate may not dilute out to end point equivalently due, for example, to differences in aggregation state.…”

Section: Results A22l Prp Sc Purification and Infectivitymentioning

confidence: 53%

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Cryo-EM of prion strains from the same genotype of host identifies conformational determinants

Hoyt

Alam

Artikis

et al. 2022

Preprint

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Prion strains in a given type of mammalian host are distinguished by differences in clinical presentation, neuropathological lesions, survival time, and characteristics of the infecting prion protein (PrP) assemblies. Near-atomic structures of prions from two host species with different PrP sequences have been determined but comparisons of distinct prion strains of the same amino acid sequence are needed to identify purely conformational determinants of prion strain characteristics. Here we report a 3.2 Å resolution cryogenic electron microscopy-based structure of the 22L prion strain purified from the brains of mice engineered to express only PrP lacking glycophosphatidylinositol anchors (a22L). Comparison of this near-atomic structure to our recently determined structure of the aRML strain propagated in the same inbred mouse reveals that these two mouse prion strains have distinct conformational templates for growth via incorporation of PrP molecules of the same sequence. Both a22L and aRML are assembled as stacks of PrP molecules forming parallel in-register intermolecular β-sheets and intervening loops, with single monomers spanning the ordered fibril core. Each monomer shares an N-terminal steric zipper, three major β-arches, and an overall V-shape, but the details of these and other conformational features differ markedly. Thus, variations in shared conformational motifs within a parallel in-register β-stack fibril architecture provide a structural basis for prion strain differentiation within a single host genotype.

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Section: Results A22l Prp Sc Purification and Infectivitymentioning

confidence: 53%

“…Fast Fourier transforms of such images indicated regular spacings of ~5.0 Å (Fig. 2B, prior to pixel size correction used in 3D reconstruction below) as was observed with 263K and aRML prions (5,6). 2D class averages were obtained from a total of 4449 images (Supplemental Fig.…”

Section: Single-particle Analysis and 3d Image Reconstructionmentioning

confidence: 73%

Cryo-EM of prion strains from the same genotype of host identifies conformational determinants

Hoyt

Alam

Artikis

et al. 2022

Preprint

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“…1a and c, d). Proteinase K (PK)digestion of PrP Sc generates a classical PrP 27-30 banding pattern on SDS-PAGE gels/western blots, comprising C-terminal proteolytic fragments of di-, mono-, and nonglycosylated PrP (Meyer et al 1986;Prusiner 1998;Wenborn et al 2015;Kraus et al 2021;Manka et al 2022b;Hoyt et al 2022). The overall similarity of PrP 27-30 banding patterns seen across multiple human and animal prion strains is indicative of a generic fibrillar prion architecture.…”

Section: Introductionmentioning

confidence: 99%

Prion strains viewed through the lens of cryo-EM

et al. 2022

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Mammalian prions are lethal transmissible pathogens that cause fatal neurodegenerative diseases in humans and animals. They consist of fibrils of misfolded, host-encoded prion protein (PrP) which propagate through templated protein polymerisation. Prion strains produce distinct clinicopathological phenotypes in the same host and appear to be encoded by distinct misfolded PrP conformations and assembly states. Despite fundamental advances in our understanding of prion biology, key knowledge gaps remain. These include precise delineation of prion replication mechanisms, detailed explanation of the molecular basis of prion strains and inter-species transmission barriers, and the structural definition of neurotoxic PrP species. Central to addressing these questions is the determination of prion structure. While high-resolution definition of ex vivo prion fibrils once seemed unlikely, recent advances in cryo-electron microscopy (cryo-EM) and computational methods for 3D reconstruction of amyloids have now made this possible. Recently, near-atomic resolution structures of highly infectious, ex vivo prion fibrils from hamster 263K and mouse RML prion strains were reported. The fibrils have a comparable parallel in-register intermolecular β-sheet (PIRIBS) architecture that now provides a structural foundation for understanding prion strain diversity in mammals. Here, we review these new findings and discuss directions for future research.

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“…Moreover, determining the high resolution structural properties of PrP Sc produced by infection with different prion strains offered an opportunity to finally elucidate the details by which heritable information is enciphered by distinct prion conformations. The two current studies in Nature Communications by Hoyt et al and Manka et al shed light on these issues 10 , 11 . Both studies involve cryo-EM analyses of the widely studied mouse-adapted scrapie isolate referred to as RML.…”

Section: Determining the Structure Of Infectious Prionsmentioning

confidence: 93%

The shape of things to come: structural insights into how prion proteins encipher heritable information

Telling

2022

Nat Commun

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The prion hypothesis embodies the radical concept that prion proteins contain the necessary information for infectious replication within their shape, thus obviating the requirement for genomic material. Two elegant papers by Hoyt et al. and Manka et al. describing high-resolution structures of infectious prions bring us closer to answering the long-standing question of how different prion conformations produce heritably distinct diseases.

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Cryo-EM structure of anchorless RML prion reveals variations in shared motifs between distinct strains

Cited by 61 publications

References 37 publications

Cryo-EM of prion strains from the same genotype of host identifies conformational determinants

Cryo-EM of prion strains from the same genotype of host identifies conformational determinants

Prion strains viewed through the lens of cryo-EM

The shape of things to come: structural insights into how prion proteins encipher heritable information

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