Cytomegalovirus Downregulates IRE1 to Repress the Unfolded Protein Response

Abstract

During viral infection, a massive demand for viral glycoproteins can overwhelm the capacity of the protein folding and quality control machinery, leading to an accumulation of unfolded proteins in the endoplasmic reticulum (ER). To restore ER homeostasis, cells initiate the unfolded protein response (UPR) by activating three ER-to-nucleus signaling pathways, of which the inositol-requiring enzyme 1 (IRE1)-dependent pathway is the most conserved. To reduce ER stress, the UPR decreases protein synthesis, increases degradation of unfolded proteins, and upregulates chaperone expression to enhance protein folding. Cytomegaloviruses, as other viral pathogens, modulate the UPR to their own advantage. However, the molecular mechanisms and the viral proteins responsible for UPR modulation remained to be identified. In this study, we investigated the modulation of IRE1 signaling by murine cytomegalovirus (MCMV) and found that IRE1-mediated mRNA splicing and expression of the X-box binding protein 1 (XBP1) is repressed in infected cells. By affinity purification, we identified the viral M50 protein as an IRE1-interacting protein. M50 expression in transfected or MCMV-infected cells induced a substantial downregulation of IRE1 protein levels. The N-terminal conserved region of M50 was found to be required for interaction with and downregulation of IRE1. Moreover, UL50, the human cytomegalovirus (HCMV) homolog of M50, affected IRE1 in the same way. Thus we concluded that IRE1 downregulation represents a previously undescribed viral strategy to curb the UPR.

Author Summary

Viruses abuse the cell's protein synthesis and folding machinery to produce large amounts of viral proteins. This enforced synthesis overloads the cell's capacity and leads to an accumulation of unfolded proteins in the endoplasmic reticulum (ER) resulting in ER stress, which can compromise cell viability. To restore ER homeostasis, cells initiate the unfolded protein response (UPR) to reduce protein synthesis, increase degradation of unfolded proteins, and upregulate chaperone expression for enhanced protein folding. The most conserved branch of the UPR is the signaling pathway activated by the ER stress sensor IRE1. It upregulates ER-associated degradation (ERAD), thereby antagonizing ER stress. Some of the counter-regulatory mechanisms of the UPR are detrimental for viral replication and are, therefore, moderated by viruses. In this study we identified the first viral IRE1 inhibitor: The murine cytomegalovirus M50 protein, which interacts with IRE1 and induces its degradation. By this means, M50 inhibits IRE1 signaling and prevents ERAD upregulation. Interestingly, the M50 homolog in human cytomegalovirus, UL50, also downregulated IRE1 revealing a previously unknown mechanism of viral host cell manipulation.