The essential yeast nuclear pore protein NSP1 was placed under the control of the regulatable GAL10 promoter. GAL::NSP1 cells grow normally in galactose medium, but arrest in growth upon glucose-induced repression of the GAL::nsp1 gene. During NSP1 depletion, nuclear accumulation of two reporter proteins Mat alpha 2-lacZ and PHO2-lacZ is inhibited, and the chimeric proteins appear in the cytoplasm of GAL::nsp1 cells. Furthermore, the nuclear pore density decreases within the nuclear membrane during early NSP1 depletion. Upon reinduction of the NSP1 gene after NSP1 depletion, NSP1 is targeted to the nuclear envelope, the nuclear pore density increases, and nuclear accumulation of reporter proteins is restored.
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PLoS Pathog
February 2024
Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, California, United States of America.
A better mechanistic understanding of virus-host dependencies can help reveal vulnerabilities and identify opportunities for therapeutic intervention. Of particular interest are essential interactions that enable production of viral proteins, as those could target an early step in the virus lifecycle. Here, we use subcellular proteomics, ribosome profiling analyses and reporter assays to detect changes in protein synthesis dynamics during SARS-CoV-2 (CoV2) infection.
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December 2023
Department of Biology, Indiana University, Bloomington, Indiana, USA.
Rotavirus is an enteric RNA virus that causes severe dehydrating gastroenteritis in infants and young children through infection of enterocytes in the small intestine. Timely clearance of the virus demands a robust innate immune response by cells associated with the small intestine, including the expression of interferon (IFN). Previous studies have shown that some rotavirus strains suppress the production of interferon, by inducing the degradation of mitochondrial antiviral signaling (MAVS) protein and interferon regulatory factor-3 (IRF3).
View Article and Find Full Text PDFbioRxiv
July 2023
Chan Zuckerberg Biohub - San Francisco, San Francisco, CA 94158, USA.
A better mechanistic understanding of virus-host interactions can help reveal vulnerabilities and identify opportunities for therapeutic interventions. Of particular interest are essential interactions that enable production of viral proteins, as those could target an early step in the virus lifecycle. Here, we use subcellular proteomics, ribosome profiling analyses and reporter assays to detect changes in polysome composition and protein synthesis during SARS-CoV-2 (CoV2) infection.
View Article and Find Full Text PDFPLoS Pathog
December 2022
Department of Microbiology & Immunology, Dalhousie University, Halifax, Canada.
Stress granules (SGs) are cytoplasmic condensates that often form as part of the cellular antiviral response. Despite the growing interest in understanding the interplay between SGs and other biological condensates and viral replication, the role of SG formation during coronavirus infection remains poorly understood. Several proteins from different coronaviruses have been shown to suppress SG formation upon overexpression, but there are only a handful of studies analyzing SG formation in coronavirus-infected cells.
View Article and Find Full Text PDFmBio
December 2022
Department of Biology, Indiana University, Bloomington, Indiana, USA.
The interferon (IFN)-inducible 2',5'-oligoadenylate synthetase (OAS)-RNase L pathway plays a critical role in antiviral immunity. Group A rotaviruses, including the simian SA11 strain, inhibit this pathway through two activities: an E3-ligase related activity of NSP1 that degrades proteins necessary for IFN signaling, and a phosphodiesterase (PDE) activity of VP3 that hydrolyzes the RNase L-activator 2',5'-oligoadenylate. Unexpectedly, we found that a recombinant (r) SA11 double mutant virus deficient in both activities (rSA11-VP3H797R-NSP1ΔC17) retained the ability to prevent RNase L activation.
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