Dataset Information

Human Respiratory Syncytial Virus Non-Structural Protein NS1 enters the nucleus to disrupt gene transcription (ChIP-seq)

ABSTRACT: Human respiratory syncytial virus (RSV) is a common cause of lower respiratory tract infections in young children, the elderly, and immunocompromised individuals. Prior exposure to RSV affords little protection and many are susceptible to reinfection throughout life. Virally encoded non-structural (NS) protein 1 (NS1) is thought to modulate host responses in the cytosol. Here, we describe the nuclear localization of NS1 during RSV infection and a corresponding nuclear role in modulating host transcription. In support, we show that a significant proportion of NS1 is partitions to the nucleus and that NS1 alone is necessary and sufficient to translocate into the nucleus. NS1 co-localizes with exportin XPO1 and inhibition of XPO1 results in NS1 accumulation in the nucleus. Furthermore, nuclear NS1 is chromatin-associated and co-immunoprecipitates with Mediator complex proteins. Chromatin-immunoprecipitation demonstrates enrichment of NS1 binding overlapping Mediator and interferon-stimulated transcription factor binding sites that lie within regulatory elements of genes differentially expressed during RSV infection. Mutation of the unique alpha helix in NS1 enhances its repressive effect on host gene expression. Together, these data suggest that nuclear NS1 may alter host responses to RSV infection by binding at the promoters and enhancers of host immune response genes and disrupting host transcriptional regulators. Our study identifies yet another regulatory layer of interactions with RSV proteins that shapes host response to RSV and potentially impacts long-term immunity to RSV.

ORGANISM(S): Homo sapiens

PROVIDER: GSE155150 | GEO | 2021/10/12

REPOSITORIES: GEO

ACCESS DATA

Similar Datasets

Project description:The NS1 protein of influenza A virus (IAV) is a multifunctional virulence factor. Mouse adaptive mutations in the NS1 protein of the human isolate A/Hong Kong/1/1968(H3N2) (HK) have been previously reported to increase virulence, viral fitness, and interferon antagonism, but differ in binding to post-transcriptional processing factor CPSF30. Because nuclear trafficking is a major genetic determinant of influenza virus host adaptation, we assessed subcellular localization and host gene expression of NS1 adaptive mutations. Recombinant HK viruses with adaptive mutations in the NS1 gene were assessed for NS1 protein subcellular localization in mouse and human cells using confocal microscopy and cellular fractionation. HK-wt virus NS1 partitioned equivalently between the cytoplasm and nucleus in human cells but was defective in cytoplasmic localization in mouse cells. The adaptive mutations either increased the proportion or abundance of NS1 in the cytoplasm, and/or the nucleus. NS1 mutations that increased cytoplasmic distribution identified a putative second nuclear export signal (NES) spanning aa positions 98-106 LSEDWFMLM, (mutation sites in bold); with the strongest effect seen for mutation M106I. The putative NES in the NS3 protein was associated with cytoplasmic localization. The host gene expression profile of the adaptive mutants was determined by microarray analysis of infected mouse cells to show either high or low gene regulation (HGR or LGR) phenotypes that mapped to the amino-terminal and the carboxy-terminal regions respectively. The HGR and LGR mutations were predominantly down regulating versus up regulating respectively. The greatest effect on host gene expression in the HGR group correlated with the ability of the NS1 protein to bind CPSF30. To our knowledge this is the first report of roles of adaptive NS1 mutations that affect intracellular localization and regulation of host gene expression. biological triplicates of mock infected mouse M1 cells; cells infected with A/HK/1/1968(H3N2) wt and NS1 mtuatn with mutaions, D2N, V23A, L98S, L98S + M106I, F103L, M106V, M106V + M124I, D125G, V180A, V226I, M106I, and R227K. Cells were infected at a multiplicty of infection of 2 and cels were incubated for 8 hr at 37 C for 8 hrs before RNA extraction and analysis relative to mock PBS infected cells.

Dataset Information

Human Respiratory Syncytial Virus Non-Structural Protein NS1 enters the nucleus to disrupt gene transcription (ChIP-seq)

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure