The UFM1 Pathway Impacts HCMV US2-Mediated Degradation of HLA Class I.

To prevent accumulation of misfolded proteins in the endoplasmic reticulum, chaperones perform quality control on newly translated proteins and redirect misfolded proteins to the cytosol for degradation by the ubiquitin-proteasome system. This pathway is called ER-associated protein degradation (ERAD). The human cytomegalovirus protein US2 induces accelerated ERAD of HLA class I molecules to prevent immune recognition of infected cells by CD8+ T cells. Using US2-mediated HLA-I degradation as a model for ERAD, we performed a genome-wide CRISPR/Cas9 library screen to identify novel cellular factors associated with ERAD. Besides the identification of known players such as TRC8, p97, and UBE2G2, the ubiquitin-fold modifier1 (UFM1) pathway was found to affect degradation of HLA-I. UFMylation is a post-translational modification resembling ubiquitination. Whereas we observe ubiquitination of HLA-I, no UFMylation was detected on HLA-I or several other proteins involved in degradation of HLA-I, suggesting that the UFM1 pathway impacts ERAD in a different manner than ubiquitin. Interference with the UFM1 pathway seems to specifically inhibit the ER-to-cytosol dislocation of HLA-I. In the absence of detectable UFMylation of HLA-I, UFM1 may contribute to US2-mediated HLA-I degradation by misdirecting protein sorting indirectly. Mass spectrometry analysis of US2-expressing cells showed that ribosomal proteins are a major class of proteins undergoing extensive UFMylation; the role of these changes in protein degradation may be indirect and remains to be established.

Screening for West Nile virus infections of susceptible animal species in Austria.

Avian mortality and encephalomyelitis in equines are considered good indicators for West Nile virus (WNV) activity. We retrospectively tested 385 horse sera for WNV antibodies and looked for WNV nucleic acid and/or WNV antigen in paraffin embedded tissues from 12 horses with aetiologically unresolved encephalomyelitis and 102 free-living birds of different species which had been found dead. With the exception of four horses originating from eastern European countries investigated on the occasion of transit through Austria, all horse sera were negative. Nested RT-PCR of the horse tissues yielded no amplification of WNV-RNA. Also, all bird samples, examined by immunohistochemistry, in situ hybridization and nested RT-PCR were negative for WNV. These results indicate that currently WNV cannot be considered a significant pathogen in Austria.