TY  - JOUR
A1  - Laine, Romain F.
A1  - Albecka, Anna
A1  - van de Linde, Sebastian
A1  - Rees, Eric J.
A1  - Crump, Colin M.
A1  - Kaminski, Clemens F.
T1  - Structural analysis of herpes simplex virus by optical super-resolution imaging
JF  - Nature Communications
N2  - Herpes simplex virus type-1 (HSV-1) is one of the most widespread pathogens among humans. Although the structure of HSV-1 has been extensively investigated, the precise organization of tegument and envelope proteins remains elusive. Here we use super-resolution imaging by direct stochastic optical reconstruction microscopy (dSTORM) in combination with a model-based analysis of single-molecule localization data, to determine the position of protein layers within virus particles. We resolve different protein layers within individual HSV-1 particles using multi-colour dSTORM imaging and discriminate envelope-anchored glycoproteins from tegument proteins, both in purified virions and in virions present in infected cells. Precise characterization of HSV-1 structure was achieved by particle averaging of purified viruses and model-based analysis of the radial distribution of the tegument proteins VP16, VP1/2 and pUL37, and envelope protein gD. From this data, we propose a model of the protein organization inside the tegument.
KW  - tegument protein pUL36
KW  - fluorescence microscopy
KW  - monoclonal antibodies
KW  - 3-dimensional structure
KW  - type-1
KW  - nuclear pore complex
KW  - reconstruction microscopy
KW  - localization microscopy
KW  - resolution
KW  - envelopment
Y1  - 2015
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-144623
VL  - 6
IS  - 5980
ER  - 
TY  - JOUR
A1  - Rutkowski, Andrzej J.
A1  - Erhard, Florian
A1  - L'Hernault, Anne
A1  - Bonfert, Thomas
A1  - Schilhabel, Markus
A1  - Crump, Colin
A1  - Rosenstiel, Philip
A1  - Efstathiou, Stacey
A1  - Zimmer, Ralf
A1  - Friedel, Caroline C.
A1  - Dölken, Lars
T1  - Widespread disruption of host transcription termination in HSV-1 infection
JF  - Nature Communications
N2  - 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 extensive transcription for tens of thousands of nucleotides beyond poly(A) sites and into downstream genes, leading to novel intergenic splicing between exons of neighbouring cellular genes. As a consequence, hundreds of cellular genes seem to be transcriptionally induced but are not translated. In contrast to previous reports, we show that HSV-1 does not inhibit co-transcriptional splicing. Our approach thus substantially advances our understanding of HSV-1 biology and establishes HSV-1 as a model system for studying transcription termination.
KW  - herpes simplex virus
KW  - RNA polymerase II
KW  - gene expression
KW  - alpha-globin
KW  - motif discovery
KW  - regulatory protein ICP27
KW  - poly(A) site usage
KW  - pre-messenger RNA
KW  - splicing inhibition
KW  - type 1 ICP27
Y1  - 2015
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-148643
VL  - 6
IS  - 7126
ER  -