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Wide-spread disruption of transcription termination in HSV-1 infection: Next generation sequencing of total and newly transcribed (4sU-RNA) RNA

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. Newly transcribed RNA was labelled by adding 500µM 4-thiouridine (4sU) to the cell culture media for 1h. Total cellular RNA was isolated using Trizol. Newly transcribed RNA was purified following the protocol described in Raedle et al. JoVE 2013. Newly transcribed RNA was labelled in one hour intervals during the first eight hours of HSV-1 infection. All nine 4sU-RNA samples as well as total cellular RNA of every second hour of infection were sent for sequencing. Two independent biological replicates were analysed.

ORGANISM(S): Homo sapiens

SUBMITTER: Caroline Friedel

PROVIDER: E-GEOD-59717 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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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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