Studying the RNA biology, cell biology, and immunology of RNA virus infection
Our laboratory studies the molecular mechanisms that regulate RNA virus-host interactions. We focus primarily on viruses in the Flaviviridae family of viruses, such as dengue virus (DENV), Zika virus (ZIKV), and hepatitis C virus (HCV). Our research is focused on defining (1) how these viruses replicate, (2) the mechanisms that regulate antiviral innate immunity to these viruses, and (3) RNA regulatory controls to both of these processes. Our long-term goal is to discover the necessary mechanistic and functional information to guide future development of new strategies for virus treatment and prevention.
Molecular mechanisms of antiviral innate immunity and viral evasion
Regulation of intracellular innate immunity is essential to prevent constitutive production of interferon (IFN). Intracellular innate immunity to RNA virus infection (IFN-induction) is activated by the cytosolic pattern recognition receptors RIG-I and MDA5, which sense pathogen-associated molecular patterns presented by RNA viruses to drive formation of a MAVS-signaling complex. Formation of this MAVS-signaling complex results in signal transduction that leads to the transcriptional activation of type I and type III IFN and ultimately the production of IFN-stimulated genes, which limit viral replication. Many RNA viruses, including HCV, suppress the activation of this antiviral program.
Previously, we have identified cellular proteins that relocalize to MAVS-signaling sites at ER-mitochondrial membrane contact sites at mitochondrial-associated ER membranes (MAM) during IFN-induction. Further, we have found that the HCV protease, NS3-NS4A cleaves MAVS at the MAM to block this signaling. Thus, we propose that dynamic localization of host and viral proteins to the MAM helps determine the outcome of HCV and RNA virus infection. As such, current research in the lab is focused at defining how membrane-targeted and relocalized host proteins coordinate the formation of the RIG-I/MAVS-signaling complex on the MAM response and how the HCV NS3-NS4A protease is targeted to the MAM and other subcellular compartments for innate immune regulation. This work has led us to uncover new innate immune regulators, such as RAB1B. This work has also defined specific domains and amino acids of NS3-NS4A that are essential for regulating anti-HCV immunity.
RNA regulatory controls to viral infection and the host response
The host response to RNA virus infection consists of hundreds of gene expression changes in cells that regulate viral infection. Many of these gene expression changes are antiviral and limit virus replication and spread, such as the well-known IFN-stimulated genes induced by the antiviral innate immune sensing pathways that produce IFNs (see above). Some of these genes are also pro-viral and encode proteins required to promote viral replication. A successful viral infection requires tight gene regulation to balance the induction of genes of the innate immune response with those required for viral infection. While we know many of the key protein players in the antiviral innate immune sensing pathways during viral infection, we know very little about the post-transcriptional regulation to these pathways or to the gene expression changes that occur during viral infection. Specifically, post-transcriptional RNA regulatory controls, such as base modifications and associated RNA binding proteins, likely regulate these responses during infection. Viral RNA itself can also be directly regulated by base modifications and RNA binding proteins.
Recent work from our lab has found that genomic RNAs of viruses in the Flaviviridae family, including HCV, ZIKV, DENV, yellow fever, and West Nile virus all contain the RNA base modification m6A within their RNA genomes, and we have found that in HCV, m6A negatively regulates viral genome packaging by modulation specific RNA binding protein interactions with the viral genome. Current projects in the lab include (1) defining how direct modification of the HCV RNA genome by m6A regulates the life cycle of HCV, (2) defining the molecular mechanisms by which m6A regulates cellular RNAs during viral infection and during the host response to infection, and (3) identifying new RNA binding proteins that regulate these interactions.