The VZV ORF20 RHIM forms heteromeric amyloid fibrils with ZBP1, which are dependent on the core tetrad for stable higher-order assembly. Megan Steain Max O. D. G. Baker Chi L. L. Pham Nirukshan Shanmugam Yann Gambin Emma Sierecki Brian P. McSharry Selmir Avdic Barry Slobedman Margaret Sunde Allison Abendroth 10.1371/journal.ppat.1008473.g009 https://plos.figshare.com/articles/figure/The_VZV_ORF20_RHIM_forms_heteromeric_amyloid_fibrils_with_ZBP1_which_are_dependent_on_the_core_tetrad_for_stable_higher-order_assembly_/12642483 <p>(A) Transmission electron micrograph of protein assemblies formed from combinations of Ub-ORF20<sub>1-114</sub>, Ub-ORF20<sub>1-114</sub>mut, and YPet-ZBP1<sub>170-355</sub> fusion proteins. Scale bar represents 200 nm. (B) Confocal microscope images of heterofibrils formed from a mixture of ORF20<sub>1-114</sub>-mCherry and YPet-ZBP1<sub>170-355</sub> in ThT-containing assembly buffer. Scale bar represents 20 μm. (C) The IQIG core tetrad from ORF20 is required to trap ZBP1 in insoluble, detergent-stable aggregates. SDS-AGE analysis of monomeric or assembled forms of Ub-ORF20<sub>1-114</sub>, Ub-ORF20<sub>1-114</sub>mut, and YPetZBP1<sub>170-355</sub>, or combinations thereof. Monomeric proteins were maintained in 8 M urea before electrophoresis. Assembled samples were incubated with either water or 2% SDS at room temperature for 10 min before electrophoresis. Identity and treatment of protein indicated above each sample.</p> 2020-07-10 17:39:37 Varicella zoster virus host cell death ZBP 1-driven apoptosis VZV infection VZV capsid triplex protein amyloid complexes TNF-induced necroptosis decoy amyloid assemblies ZBP 1 pathway sequestering ZBP 1 RIP Homotypic Interaction Motif host cell death inhibitor VZV ORF 20 RHIM Varicella zoster virus encodes RIPK cell death Herpesviruses ORF 20 RHIM ORF 20 RHIM forms ZBP 1-expressing HT -29 cells