Endogenous Proteolytic Cleavage of Disease-associated Prion Protein to Produce C2 Fragments Is Strongly Cell- and Tissue-dependent

Dron, Michel; Moudjou, Mohammed; Chapuis, Jérôme; Salamat, Muhammad Khalid Farooq; Bernard, Julie; Cronier, Sabrina; Langevin, Christelle; Laude, Hubert

doi:10.1074/jbc.m109.083857

Cited by 62 publications

(68 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The C2 aspect is especially notable as Watts and colleagues recently argued that Sho is degraded via an endocytic pathway that requires it to be physically trafficked with PrP Sc (10). A component of this argument was the inverse correlation between Sho levels and the PrP C2 proteolytic fragment (10), with the latter hypothesized as a stable intermediate in the lysosomal degradation of PrP Sc (44,45). However, we have shown that models for other neurodegenerative diseases with activated lysosomes lack downregulation of Sho (11).…”

Section: Figurementioning

confidence: 89%

Prion disease tempo determined by host-dependent substrate reduction

Mays¹,

Kim²,

Haldiman³

et al. 2014

J. Clin. Invest.

View full text Add to dashboard Cite

The symptoms of prion infection can take years or decades to manifest following the initial exposure. Molecular markers of prion disease include accumulation of the misfolded prion protein (PrP Sc ), which is derived from its cellular precursor (PrP C ), as well as downregulation of the PrP-like Shadoo (Sho) glycoprotein. Given the overlapping cellular environments for PrP C and Sho, we inferred that PrP C levels might also be altered as part of a host response during prion infection. Using rodent models, we found that, in addition to changes in PrP C glycosylation and proteolytic processing, net reductions in PrP C occur in a wide range of prion diseases, including sheep scrapie, human Creutzfeldt-Jakob disease, and cervid chronic wasting disease. The reduction in PrP C results in decreased prion replication, as measured by the protein misfolding cyclic amplification technique for generating PrP Sc in vitro. While PrP C downregulation is not discernible in animals with unusually short incubation periods and high PrP C expression, slowly evolving prion infections exhibit downregulation of the PrP C substrate required for new PrP Sc synthesis and as a receptor for pathogenic signaling. Our data reveal PrP C downregulation as a previously unappreciated element of disease pathogenesis that defines the extensive, presymptomatic period for many prion strains.

show abstract

Section: Figurementioning

confidence: 89%

Prion disease tempo determined by host-dependent substrate reduction

Mays¹,

Kim²,

Haldiman³

et al. 2014

J. Clin. Invest.

View full text Add to dashboard Cite

show abstract

“…Furthermore, filamentous depositions of PrP Sc were frequently observed in the white matter of patients with inherited prion diseases (Reiniger et al, 2013) and in the neuropils of the 22L prion straininfected cerebellar slices (Wolf et al, 2015). In addition, it was reported that PrP Sc accumulates predominantly as unprocessed forms in brain tissues and in the primarycultured cerebellar-granule neurons (Dron et al, 2010). Since PrP Sc detected by mAb 8D5 is expected to be a full-length PrP Sc , therefore the morphological and locational, as well as biochemical, similarities of PrP Sc stains in prion-infected CNs and brains of patients or animals affected by prion diseases suggest that the mechanism of PrP Sc formation in CNs is, at least to some extent, similar to that in neurons in the central nervous system.…”

Section: Discussionmentioning

confidence: 99%

“…To resolve the partly contradictory results regarding the site of PrP Sc formation between cell culture and ultrastructural studies, primarycultured neurons are considered to be a good ex vivo model. There are only a few reports on prion propagation in primary-cultured neurons derived from the cerebellum, striatum, and cerebral cortex of mouse brains (Cronier et al, 2004;Dron et al, 2010;Hannaoui et al, 2013). However, little information is available on the localization of PrP Sc in primary neurons infected with prions.…”

Section: Introductionmentioning

confidence: 99%

Comparison of abnormal isoform of prion protein in prion-infected cell lines and primary-cultured neurons by PrPSc-specific immunostaining

Tanaka

Fujiwara

Suzuki

et al. 2016

Journal of General Virology

View full text Add to dashboard Cite

We established abnormal isoform of prion protein (PrP Sc )-specific double immunostaining using mAb 132, which recognizes aa 119-127 of the PrP molecule, and novel PrP Sc -specific mAb 8D5, which recognizes the N-terminal region of the PrP molecule. Using the PrP Sc -specific double immunostaining, we analysed PrP Sc in immortalized neuronal cell lines and primary cerebralneuronal cultures infected with prions. The PrP Sc -specific double immunostaining showed the existence of PrP Sc positive for both mAbs 132 and 8D5, as well as those positive only for either mAb 132 or mAb 8D5. This indicated that double immunostaining detects a greater number of PrP Sc species than single immunostaining. Double immunostaining revealed cell-type-dependent differences in PrP Sc staining patterns. In the 22 L prion strain-infected Neuro2a (N2a)-3 cells, a subclone of N2a neuroblastoma cell line, or GT1-7, a subclone of the GT1 hypothalamic neuronal cell line, granular PrP Sc stains were observed at the perinuclear regions and cytoplasm, whereas unique string-like PrP Sc stains were predominantly observed on the surface of the 22 L straininfected primary cerebral neurons. Only 14 % of PrP Sc in the 22 L strain-infected N2a-3 cells were positive for mAb 8D5, indicating that most of the PrP Sc in N2a-3 lack the N-terminal portion. In contrast, nearly half PrP Sc detected in the 22 L strain-infected primary cerebral neurons were positive for mAb 8D5, suggesting the abundance of full-length PrP Sc that possesses the Nterminal portion of PrP. Further analysis of prion-infected primary neurons using PrP Sc -specific immunostaining will reveal the neuron-specific mechanism for prion propagation.

show abstract

“…For PrP res , otherwise-indicated samples corresponding to the PK-resistant PrP present in either 25 or 50 g of protein of cellular lysate were loaded on the gel. The transfer of proteins, their detection, and their revelation were described previously (39,47).…”

Section: Methodsmentioning

confidence: 99%

Generating Bona Fide Mammalian Prions with Internal Deletions

Muñoz-Montesino

Sizun

Moudjou

et al. 2016

J Virol

Self Cite

View full text Add to dashboard Cite

Mammalian prions are PrP proteins with altered structures causing transmissible fatal neurodegenerative diseases. They are self-perpetuating through formation of beta-sheet-rich assemblies that seed conformational change of cellular PrP. Pathological PrP usually forms an insoluble protease-resistant core exhibiting beta-sheet structures but no more alpha-helical content, loosing the three alpha-helices contained in the correctly folded PrP. The lack of a high-resolution prion structure makes it difficult to understand the dynamics of conversion and to identify elements of the protein involved in this process. To determine whether completeness of residues within the protease-resistant domain is required for prions, we performed serial deletions in the helix H2 C terminus of ovine PrP, since this region has previously shown some tolerance to sequence changes without preventing prion replication. Deletions of either four or five residues essentially preserved the overall PrP structure and mutant PrP expressed in RK13 cells were efficiently converted into bona fide prions upon challenge by three different prion strains. Remarkably, deletions in PrP facilitated the replication of two strains that otherwise do not replicate in this cellular context. Prions with internal deletion were self-propagating and de novo infectious for naive homologous and wild-type PrP-expressing cells. Moreover, they caused transmissible spongiform encephalopathies in mice, with similar biochemical signatures and neuropathologies other than the original strains. Prion convertibility and transfer of strain-specific information are thus preserved despite shortening of an alpha-helix in PrP and removal of residues within prions. These findings provide new insights into sequence/structure/infectivity relationship for prions. IMPORTANCEPrions are misfolded PrP proteins that convert the normal protein into a replicate of their own abnormal form. They are responsible for invariably fatal neurodegenerative disorders. Other aggregation-prone proteins appear to have a prion-like mode of expansion in brains, such as in Alzheimer's or Parkinson's diseases. To date, the resolution of prion structure remains elusive. Thus, to genetically define the landscape of regions critical for prion conversion, we tested the effect of short deletions. We found that, surprisingly, removal of a portion of PrP, the C terminus of alpha-helix H2, did not hamper prion formation but generated infectious agents with an internal deletion that showed characteristics essentially similar to those of original infecting strains. Thus, we demonstrate that completeness of the residues inside prions is not necessary for maintaining infectivity and the main strain-specific information, while reporting one of the few if not the only bona fide prions with an internal deletion.

show abstract

Endogenous Proteolytic Cleavage of Disease-associated Prion Protein to Produce C2 Fragments Is Strongly Cell- and Tissue-dependent

Cited by 62 publications

References 61 publications

Prion disease tempo determined by host-dependent substrate reduction

Prion disease tempo determined by host-dependent substrate reduction

Comparison of abnormal isoform of prion protein in prion-infected cell lines and primary-cultured neurons by PrPSc-specific immunostaining

Generating Bona Fide Mammalian Prions with Internal Deletions

Contact Info

Product

Resources

About