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RNA sequencing of HTNV NP and Gc proteins expressing cells

ABSTRACT: RNA sequencing of HTNV NP and Gc proteins expressing cells

PROVIDER: PRJNA966084 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR24391863_1.fastq.gz	Fastqsanger.gz
	SRR24391863_2.fastq.gz	Fastqsanger.gz
	SRR24391864_1.fastq.gz	Fastqsanger.gz
	SRR24391864_2.fastq.gz	Fastqsanger.gz
	SRR24391865_1.fastq.gz	Fastqsanger.gz

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RNA sequencing of HTNV NP and Gc proteins expressing cells

Project description:Hantavirus infection has become a public health concern worldwide due to its ability to cause severe diseases in humans. The virulence of Hantavirus is determined by their ability to restrain host innate immune responses, such as type Ⅰ interferon (IFN-Ⅰ) response. Here, we demonstrated that Hantaan virus (HTNV), a prevalent pathogen that causes hemorrhagic fever with renal syndrome in China, can inhibit host IFN-Ⅰ production by manipulating the retinoic acid-induced gene I (RIG-I)-like receptor (RLRs) pathways. In brief, we performed the transcriptome analysis to identify the host factors regulated by nucleocapsid protein (NP) and glycoprotein (Gc) of HTNV, which showed that multiple cellular biological pathways were regulated by NP and Gc, including interferon signaling pathways, neuron differentiation, and response to mechanical stimulus. Further analysis demonstrated that HTNV NP and Gc proteins inhibited IFN-Ⅰ response by targeting MAVS upstream molecules, such as RIG-I and MDA-5. Mechanistically, NP and Gc proteins suppressed RIG-I and MDA-5-induced IFN-Ⅰ production by targeting TRIM25, rather than directly actin on RIG-I and MDA-5. Taken together, our study revealed a cross-talk between HTNV NP or Gc proteins and host innate immune response, which is helpful in the understanding of virus-host interaction in general.

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Project description:The oncolytic potential of NDV has gained significant attention in the context of clinical trials. After systematically comparing the oncolytic activities of different NDV subtypes,we found that all subtypes, except for highly pathogenic genotype VII NDVs, display remarkable infectivity on tumor cell lines.A single amino acid mutation (F450L) in the NP protein at position 450 leads to an approximate 1000000-fold increase in the TCID50 value of genotype VII NDVs on tumor cells.Mechanistic inquiries unveiled that the amino acid at position 450 of the NP protein governs preferential translation of NDV mRNA.The preferential translation that regulates viral mRNA translation is often accompanied by modulation of the host translation machinery. Therefore, we established HeLa cell lines stably expressing NP protein from different NDV strains. Ribosome-bound mRNA was enriched to perform Ribosome Nascent-chain Complex sequencing(RNC-seq), while total cellular mRNA was enriched to conducted RNA-Seq . Untreated cells were used as controls. These experiments aimed to investigate the impact of NP on the host translation system.

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RNA-sequencing of pre-GC B cells helped by CD4-CrexBcl6fl/+ or CD4-Cre OT-II T cells

Project description:We analyzed transcriptome differences of antigen-specific pre-GC B cells. We transferred wildtype (CD4-Cre) or BCL6-insufficient (CD4-CrexBcl6fl/+) OT-II T cells into SAP-deficient mice and immunized these mice with NP-OVA. By day 3 post immunization, we sort-purified NP-binding B cells that were developing into GCs and conducted RNA-seq analyses of their transcriptome. NP+IgDlowB220+ pre-GC B cells from recipients of the same group were pooled to sort-purify 4 200-cell repeats. Using a gene expression signature induced by CD40 signaling in B cells for enrichment analysis, we found such CD40-induced gene set expression was significantly deprived in those NP-binding B cells helped by CD4-Cre´Bcl6fl/+ T cells. These data suggest BCL6 insufficiency in T cells leads to impaired CD40L signaling to B cells.

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