Identification of NLR-associated Amyloid Signaling Motifs in Bacterial Genomes

• Published in: Journal of molecular biology 2020-11, Vol.432 (23), p.6005-6027
• Main Authors: Dyrka, Witold, Coustou, Virginie, Daskalov, Asen, Lends, Alons, Bardin, Thierry, Berbon, Mélanie, Kauffmann, Brice, Blancard, Corinne, Salin, Bénédicte, Loquet, Antoine, Saupe, Sven J.
• Format: Article
• Language: English
• Subjects: actinobacteria; Amino Acid Motifs - genetics; Amino Acid Sequence - genetics; Amyloid - genetics; Amyloidogenic Proteins - genetics; Animals; cyanobacteria; Cyanobacteria - genetics; Drosophila - genetics; Evolution, Molecular; Fungi - genetics; Genome, Bacterial - genetics; Immunity, Innate - genetics; Life Sciences; multicellularity; NLR Proteins - genetics; Prions - genetics; programmed cell death; signal transduction; Signal Transduction - genetics
• ISSN: 0022-2836; 1089-8638
• DOI: 10.1016/j.jmb.2020.10.004

[Display omitted] •Bacterial genomes contain amyloid motifs associated to homologs of Nod-like receptors.•Bacterial amyloid signaling sequences (BASS) occur mainly in multicellular bacteria.•Selected BASS-motifs propagate as prions in a fungal model.•BASS3 motifs cross-seed in vivo with RHIM and fungal PP-motifs. In filamentous fungi, amyloid signaling sequences allow Nod-like receptors (NLRs) to activate downstream cell-death inducing proteins with HeLo and HeLo-like (HELL) domains and amyloid RHIM and RHIM-related motifs control immune defense pathways in mammals and flies. Herein, we show bioinformatically that analogous amyloid signaling motifs exist in bacteria. These short motifs are found at the N terminus of NLRs and at the C terminus of proteins with a domain we term BELL. The corresponding NLR and BELL proteins are encoded by adjacent genes. We identify 10 families of such bacterial amyloid signaling sequences (BASS), one of which (BASS3) is homologous to RHIM and a fungal amyloid motif termed PP. BASS motifs occur nearly exclusively in bacteria forming multicellular structures (mainly in Actinobacteria and Cyanobacteria). We analyze experimentally a subset of seven of these motifs (from the most common BASS1 family and the RHIM-related BASS3 family) and find that these sequences form fibrils in vitro. Using a fungal in vivo model, we show that all tested BASS-motifs form prions and that the NLR-side motifs seed prion-formation of the corresponding BELL-side motif. We find that BASS3 motifs show partial prion cross-seeding with mammalian RHIM and fungal PP-motifs and that proline mutations on key positions of the BASS3 core motif, conserved in RHIM and PP-motifs, abolish prion formation. This work expands the paradigm of prion amyloid signaling to multicellular prokaryotes and suggests a long-term evolutionary conservation of these motifs from bacteria, to fungi and animals.