Prion cross-seeding between fungal and mammalian amyloid signaling motifs

Abstract

In filamentous fungi, NLR-based signalosomes activate downstream membrane-targeting cell-death inducing proteins by a mechanism of amyloid templating. In the species Podospora anserina, two such signalosomes, NWD2/HET-S and FNT1/HELLF have been described. An analogous system, involving a distinct amyloid signaling motif termed PP was also identified in the genome of the species Chaetomium globosum and studied using heterologous expression in Podospora anserina. The PP-motif bears resemblance to the RHIM and RHIM-like motifs controlling necroptosis in mammals and innate immunity in flies. We identified here, a third NLR signalosome in Podospora anserina comprising a PP-motif and organized as a two-gene cluster encoding a NLR and a HELL-domain cell-death execution protein termed HELLP. We show that the PP-motif region of HELLP forms a prion we term [{pi}] and that [{pi}] prions trigger the cell-death inducing activity of full length HELLP. We detect no prion cross-seeding between HET-S, HELLF and HELLP amyloid motifs. In addition, we find that akin to PP-motifs, RHIM motifs from human RIP1 and RIP3 kinases are able to form prions in Podospora, and that [{pi}] and [Rhim] prions partially cross-seed. Our study shows that Podospora anserina displays three independent cell-death inducing amyloid signalosomes. Based on the described functional similarity between RHIM and PP, it appears likely that these amyloid motifs constitute evolutionary related cell-death signaling modules. ImportanceAmyloids are {beta}-sheet-rich protein polymers that can be pathological or display a variety of biological roles. In filamentous fungi, specific immune receptors activate programmed cell-death execution proteins through a process of amyloid templating akin to prion propagation. Among these fungal amyloid signaling sequences, the PP-motif stands out because it shows similarity to RHIM, an amyloid sequence controlling necroptotic cell-death in mammals. We characterized an amyloid signaling system comprising a PP-motif in the model species Podospora anserina thus bringing to three the number of independent amyloid signaling cell death pathways described in that species. We then show that human RHIM motifs not only propagate as prions in P. anserina but also partially cross-seed with fungal PP-prions. These results indicate that in addition to show sequence similarity, PP and RHIM-motif are at least partially functionally related, supporting a model of long-term evolutionary conservation of amyloid signaling mechanisms from fungi to mammals.