Biology subjects

David, C. T.

Publications and source records attributed to David, C. T..

3 recordsLinked to original sources

bioRxiv2026

The rabies virus interferon antagonist P protein selectively modulates interferon signalling to inhibit antiviral gene expression while supporting proviral gene expression

Type-I IFNs mediate the principle antiviral response of cells by controlling the expression of hundreds of IFN-regulated genes (IRGs), many of which have antiviral functions. The best understood mediators of IFN signalling are STAT1 and STAT2, and STAT1/2-dependent gene induction is conventionally viewed as the primary outcome of type-I IFN signalling. To overcome the IFN response, viruses express proteins called IFN-antagonists, which target IFN signalling pathways (e.g. rabies virus P-protein (RABV-P) binds and inhibits IFN-activated STAT1/2) and so are typically considered to mediate shutdown of the IFN response. However, IFN signalling is not exclusively antiviral, with many IRGs reported to be required for or to facilitate infection by certain viruses. How viruses coordinate the apparent need to suppress certain antiviral IRGs, while presumably permitting the expression of others, including proviral IRGs is poorly defined. However, it has been shown that type-I IFN can activate multiple pathways other than classical STAT1/2, so discriminatory targeting of specific pathways by IFN-antagonists may enable highly selective regulation of distinct IRGs, dependent on the requirements of the specific virus. Here, we analyse the global effects of RABV-P protein on the IFN-regulated transcriptome. We confirm that IFN not only stimulates (IFN-stimulated genes, ISGs) but also represses (IFN-repressed genes, IRepGs) a large number of IRGs. Notably, our data indicate that RABV-P protein can antagonize both the IFN-dependent stimulation and repression of certain ISGs and IRepGs, without significantly impacting the expression of large proportion of IRGs. Antagonized ISGs included classical antiviral genes, while non-antagonized ISGs include genes with pro-viral effects on RABV. Transcription factor analysis indicated that RABV-P antagonizes STAT1/2-regulated IRGs, but not IRGs regulated by other pathways including MAP-kinase pathways, which are important to process such as cell survival and inflammatory response. These data indicate that selective modulation rather than global inhibition of IFN-signalling has beneficial outcomes for replication. The data also support the significance of IRepGs, and their modulation by IFN antagonists in viral infection. Significance StatementThe ability of viruses to evade immunity is critical to disease and so presents targets for the development of interventions. The principle antiviral response of cells is mediated by interferons (IFNs), which activate STAT proteins to induce hundreds of genes including antiviral genes. Viruses counter this by expressing IFN-antagonist proteins, many of which directly inhibit STATs. IFN-antagonists are typically considered to shut down IFN responses, but many IRGs have pro-viral functions. How viruses coordinate the apparent need to antagonise some IRGs but not others are poorly understood. Using a well-characterised viral IFN-antagonist, we find that by selectively targeting certain IFN-activated pathways, IFN-antagonists can inhibit effects of IFN on specific subsets of IRGs (including antiviral genes) without affecting others (including proviral genes); thus, IFN-antagonists may be redefined as selective IFN-modulators.

systems biology↗

bioRxiv2026

Rabies virus antagonizes interferon signaling by targeting phosphorylated STAT1 tetramers

AO_SCPLOWBSTRACTC_SCPLOWRabies virus phosphoprotein (P protein) antagonizes interferon signalling by selectively targeting activated STAT1 protein, but the structural basis for the selective recognition of activated STAT1 has remained unclear. Here we report cryo-EM structures of tyrosine-phosphorylated STAT1 (pY-STAT1) in complex with the C-terminal domain of P protein (P-CTD). P-CTD engages the pY-STAT1 tetramer through three discrete interfaces, contacting two DNA-binding domains within a pY-mediated STAT1 dimer and the N-terminal domain of a third STAT1 protomer. This binding mode explains how P-CTD selectively recognizes activated STAT1, occludes surfaces required for importin 5 and DNA binding, and stabilizes the compact tetrameric state, thereby restricting the conformational transition required for cooperative DNA binding. Cell-based assays further indicate that distinct P-CTD surfaces differentially contribute to antagonism of type I and type II interferon signalling. Further, by acting as an anchoring molecule, P-CTD also enabled structural elucidation of the complete STAT1 structures in the tetramer. Together, these data reveal critical mechanisms in viral immune evasion and pathogenesis, and fundamental innate immune signaling.

molecular biology↗

bioRxiv2023

Sub-nucleolar trafficking of Hendra virus matrix protein is regulated by ubiquitination and oligomerisation

Hendra virus (HeV) is a highly pathogenic member of the Henipavirus genus (order Mononegavirales), the replication cycle of which occurs primarily in the cytoplasm. The HeV matrix protein (HeV M) plays critical roles in viral assembly and budding at the plasma membrane, but also undergoes nuclear/nucleolar trafficking, to accumulate in nucleoli early in infection and, later, localise predominantly at the plasma membrane. Previously we found that HeV M protein targets specific sub-nucleolar compartments (corresponding to the FC-DFC (fibrillar centre (FC)/dense fibrillar component (DFC)) where it interacts with the nucleolar protein Treacle and modulates rRNA biogenesis by subverting the host nucleolar DNA damage response, indicating the importance of specific sub-nucleolar trafficking to infection. However, the mechanisms underlying targeting and movement between sub-nucleolar compartments by viral or cellular proteins remain poorly defined. Here, we assessed the molecular regulation of HeV M protein nucleolar/sub-nucleolar trafficking, finding that in infected cells and in cells expressing HeV M protein alone, M protein localizes into Treacle-enriched FC-DFC at early time points, and that FC-DFC localization is subsequently lost due to relocalization into the surrounding granular component (GC) of the nucleolus. Analysis using mutated M proteins and pharmacological modulation of ubiquitination indicate that this dynamic localization is regulated by ubiquitination and oligomerisation, with ubiquitination required for retention of HeV M in Treacle-enriched sub-nucleolar compartments, and oligomerisation required for egress. To our knowledge, this study provides the first direct insights into the dynamics and mechanisms of viral protein trafficking between sub-nucleolar compartments, important to the interplay between HeV M protein and host cell factors during infection. AUTHOR SUMMARYHenipaviruses, including Hendra (HeV) and Nipah viruses, cause deadly diseases in humans and livestock and are considered priority diseases by the World Health Organization due to their epidemic potential and lack of effective treatments. Understanding how these viruses interact with host cells is essential for developing new therapeutics. Our study examines the matrix (M) protein of henipaviruses and its interaction with the nucleolus, a cell structure that mediates ribosome production, and is a common target for various viruses, although their functions are largely unresolved. Previously, we showed that the HeV M protein targets a sub-nucleolar structure, called the FC-DFC, to modulate ribosome biogenesis. Here, we report that the M proteins movement between sub-nucleolar compartments is controlled by two processes: ubiquitination, which causes accumulation of the protein in the FC-DFC, and oligomerization, which is associated with exit. Similar mechanisms are also observed in other henipaviruses. Our findings reveal mechanisms regulating the hijacking of host cell functions by henipaviruses and suggest new potential targets for antiviral therapies. This study is the first to investigate how viral proteins move within the nucleolus, offering new insights into interactions that may be significant to multiple viruses.

microbiology↗