Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Wide-spread disruption of transcription termination in HSV-1 infection: Next-generation sequencing of translational activityd by ribosome profiling

ABSTRACT: Primary human foreskin fibroblasts (HFF) were infected with wild-type simplex virus 1 (HSV-1) strain 17 at a multiplicity of infection (MOI) of 10. Ribosome profiling was performed at various times during infection with minor modification to the protocol described in Stern-Ginossar N et al., Science 2012 Ribosome profiling was performed a 0, 1, 2, 4, 6 and 8 h post infection. Two biological replicates were analysed.

ORGANISM(S): Homo sapiens

SUBMITTER: Caroline Friedel

PROVIDER: E-GEOD-60040 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Widespread disruption of host transcription termination in HSV-1 infection.

Rutkowski Andrzej J AJ Erhard Florian F L'Hernault Anne A Bonfert Thomas T Schilhabel Markus M Crump Colin C Rosenstiel Philip P Efstathiou Stacey S Zimmer Ralf R Friedel Caroline C CC Dölken Lars L

Nature communications 20150520

Herpes simplex virus 1 (HSV-1) is an important human pathogen and a paradigm for virus-induced host shut-off. Here we show that global changes in transcription and RNA processing and their impact on translation can be analysed in a single experimental setting by applying 4sU-tagging of newly transcribed RNA and ribosome profiling to lytic HSV-1 infection. Unexpectedly, we find that HSV-1 triggers the disruption of transcription termination of cellular, but not viral, genes. This results in exten ...[more]

PMID: 25989971

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Project description:Annotating Low Abundance and Transient RNAs in Yeast using Tiling Microarrays and Ultra High-throughput Sequencing Reveals New Transcripts that are not Conserved Across Closely Related Yeast Species A complete description of the transcriptome of an organism is crucial for a comprehensive understanding of how it functions, how its transcriptional networks are controlled, and may provide insights into the organism’s evolution. Despite the status of Saccharomyces cerevisiae as arguably the most well studied model eukaryote, we still do not have a full catalog or understanding of all its genes. In order to interrogate the transcriptome of S. cerevisiae for low abundance or rapidly turned over transcripts, we deleted elements of the RNA degradation machinery with the goal of preferentially increasing the relative abundance of such transcripts. We then used high-resolution tiling microarrays and ultra high-throughput sequencing (UHTS) to identify, map and validate unannotated transcripts that are more abundant in the RNA degradation mutants relative to wild-type cells. We identified 365 currently unannotated transcripts, the majority presumably representing low abundance or short-lived RNAs, of which 185 are previously unknown and unique to this study. It is likely that many of these are cryptic unstable transcripts (CUTs), which are rapidly degraded and whose function(s) within the cell are still unclear, while others may be novel functional transcripts. Of the 185 transcripts we identified as novel to our study, greater than 80 percent come from regions of the genome that have lower conservation scores amongst closely related yeast species than 85 percent of the verified ORFs in S. cerevisiae. Such regions of the genome have typically been less well studied, and by definition transcripts from these regions will distinguish S. cerevisiae from these closely related species. Keywords: Saccharomyces cerevisiae, transcriptome, RNA-Seq, RNA degradation, XRN1, RRP6, LSM1, PAT1, Salt Shock, NaCl Four samples, a wild-type reference and three mutants (1 containing 4 deletions and the other two containing 3 each), were subjected to high salt shock, and total RNA was harvested. PolyA RNA was purified, and libraries were generated for the Solexa platform. Each library was run on 4 lanes of a Solexa flow cell.

Dataset Information

Wide-spread disruption of transcription termination in HSV-1 infection: Next-generation sequencing of translational activityd by ribosome profiling

Publications

Widespread disruption of host transcription termination in HSV-1 infection.

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