Project description:Murine gammaherpesvirus 68 (MHV-68) is closely related to Epstein-Barr virus (EBV) and Kaposi’s sarcoma-associated herpesvirus (KSHV) and provides a small animal model to study the pathogenesis of gammaherpesvirus (γHV) infections. To completely explore the potential of the MHV-68 system for the investigation of gHV miRNAs, it would be desirable to know the number and expression patterns of all miRNAs encoded by MHV-68. By using small RNA deep sequencing, we systematically investigated the MHV-68 miRNA expression profiles in both lytically and persistently infected cells. In addition to the known nine MHV-68 miRNAs, we identified six novel MHV-68 miRNA genes and analyzed the expression levels of all MHV-68 miRNAs. Furthermore, we also characterized the cellular miRNA expression signatures in MHV-68 infected versus non-infected NIH3T3 fibroblasts and in TPA-treated versus non-treated S11 cells. We found that mmu-mir-15b and mmu-mir-16 are highly upregulated upon MHV-68 infection of NIH3T3 cells, indicating a potential role of cellular miRNAs during MHV-68 infection. Our data will aid to fully explore the functions of gHV miRNAs.
Project description:Ribosome profiling (RiboSeq) (maps positions of translating ribosomes on the transcriptome) and RNASeq (quantifies the transcriptome) analysis of murine 17 clone 1 (17Cl-1) cells infected with Murine coronavirus strain A59 (MHV-A59). Samples comprise 1 and 8 h mocks, 1, 2.5, 5 and 8 h post infection timecourse, for each of RiboSeq with cycloheximide (CHX), RiboSeq with harringtonine (HAR), and RNASeq, performed in duplicate (6 x 3 x 2 libraries); RiboSeq CHX, RiboSeq HAR and RNASeq at 1 h post infection for high multiplicity of infection (3 libraries); and 1 \long-read\ library for 5 h post infection RiboSeq CHX to test for larger-than-normal ribosome footprints.
Project description:Murine gammaherpesvirus 68 (MHV-68) is closely related to Epstein-Barr virus (EBV) and KaposiM-bM-^@M-^Ys sarcoma-associated herpesvirus (KSHV) and provides a small animal model to study the pathogenesis of gammaherpesvirus (M-NM-3HV) infections. To completely explore the potential of the MHV-68 system for the investigation of gHV miRNAs, it would be desirable to know the number and expression patterns of all miRNAs encoded by MHV-68. By using small RNA deep sequencing, we systematically investigated the MHV-68 miRNA expression profiles in both lytically and persistently infected cells. In addition to the known nine MHV-68 miRNAs, we identified six novel MHV-68 miRNA genes and analyzed the expression levels of all MHV-68 miRNAs. Furthermore, we also characterized the cellular miRNA expression signatures in MHV-68 infected versus non-infected NIH3T3 fibroblasts and in TPA-treated versus non-treated S11 cells. We found that mmu-mir-15b and mmu-mir-16 are highly upregulated upon MHV-68 infection of NIH3T3 cells, indicating a potential role of cellular miRNAs during MHV-68 infection. Our data will aid to fully explore the functions of gHV miRNAs. A mouse fibroblast cell line infected with/without MHV-68 and a MHV-68 infected mouse B lymphoma cell line treated with/without TPA (4 samples in total) were examined.
Project description:In this study we explored the antiviral gene expression induced in the CNS of MHV-infected mice, by performing whole-genome expression profiling. Three different mouse strains (BALB/c, 129SvEv and 129SvEv IFNAR-/- mice), differing in their susceptibility to infection with MHV, were used.
Project description:The mouse hepatitis virus (MHV) genomic and sub-genomic RNAs are 3’ polyadenylated. The length of the 3’ poly(A) tail is known to change during infection, but little is known about the molecular mechanisms driving this change. Here, we investigate the presence of terminal uridylation and terminal guanylation on the MHV poly(A) tail during infection. We infected 17-CL1 cells with MHV and isolated RNA at 24- and 48-hour post-infection (hpi). We observed that MHV RNAs poly(A) tails shortened during infection and that short poly(A) tails of about 20 nucleotides are highly uridylated.
Project description:The mouse hepatitis virus (MHV) genomic and sub-genomic RNAs have 3’ poly(A) tails. The terminal addition of uridines to poly(A) tails has been shown to be a widespread modification. Here, we investigated the presence of 3' end additions on the MHV RNA poly(A) tails. To this end, we infected NCTC cells with MHV and isolated RNA at 24-hours post-infection (hpi). While the median poly(A) tail length of the MHV RNAs is around 50 nucleotides, we observed a peak of uridylation in transcripts with poly(A) tails about 40 nucleotides long