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bioRxiv · 10.1101/2021.02.16.431411

Human cytomegalovirus blocks canonical TGFβ signaling during lytic infection to limit induction of type I interferons

Pham, A. H.·Mitchell, J.·Botto, S.·Pryke, K. M.·Defilippis, V. R.·Hancock, M.
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Abstract

Human cytomegalovirus (HCMV) microRNAs (miRNAs) significantly rewire host signaling pathways to support the viral lifecycle and regulate host cell responses. Here we show that SMAD3 expression is regulated by HCMV miR-UL22A and contributes to the IRF7-mediated induction of type I IFNs and IFN-stimulated genes (ISGs) in human fibroblasts. Addition of exogenous TGF{beta} interferes with the replication of a miR-UL22A mutant virus in a SMAD3-dependent manner in wild type fibroblasts, but not in cells lacking IRF7, indicating that downregulation of SMAD3 expression to limit IFN induction is important for efficient lytic replication. These findings uncover a novel interplay between SMAD3 and innate immunity during HCMV infection and highlight the role of viral miRNAs in modulating these responses. Author SummaryCells trigger the interferon (IFN) response to induce the expression of cellular genes that limit virus replication. In turn, viruses have evolved numerous countermeasures to avoid the effects of IFN signaling. Using a microRNA (miRNA) mutant virus we have uncovered a novel means of regulating the IFN response during human cytomegalovirus (HCMV) infection. Lytic HCMV infection induces the production of TGF{beta}, which binds to the TGF{beta} receptor and activates the receptor-associated SMAD SMAD3. SMAD3, together with IRF7, induces the expression of IFN{beta} and downstream IFN-stimulated genes in human fibroblasts. To counteract this, HCMV miR-UL22A, along with other HCMV gene products, directly targets SMAD3 for downregulation. Infection of fibroblasts with a miR-UL22A mutant virus results in enhanced type I IFN production in a SMAD3-dependent manner and the virus is impaired for growth in the presence of TGF{beta}, but only when both SMAD3 and IRF7 are present, highlighting the unique interaction between TGF{beta} and innate immune signaling.

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Pham, A. H., Mitchell, J., Botto, S., Pryke, K. M., Defilippis, V. R., Hancock, M.. 2021-02-16. Human cytomegalovirus blocks canonical TGFβ signaling during lytic infection to limit induction of type I interferons. https://doi.org/10.1101/2021.02.16.431411

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