A loss of function analysis of host factors influencing Vaccinia virus replication by RNA interference

Griffiths, Samantha J., Beard, Philippa M., Gonzalez, Orland, Haga, Ismar R., Pechenick Jowers, Tali, Reynolds, Danielle K., Wildenhain, Jan, Tekotte, Hille, Auer, Manfred, Tyers, Mike, Ghazal, Peter ORCID: https://orcid.org/0000-0003-0035-2228, Zimmer, Ralf and Haas, Jürgen 2014. A loss of function analysis of host factors influencing Vaccinia virus replication by RNA interference. PLoS ONE 9 (6) , e98431. 10.1371/journal.pone.0098431

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Abstract

Vaccinia virus (VACV) is a large, cytoplasmic, double-stranded DNA virus that requires complex interactions with host proteins in order to replicate. To explore these interactions a functional high throughput small interfering RNA (siRNA) screen targeting 6719 druggable cellular genes was undertaken to identify host factors (HF) influencing the replication and spread of an eGFP-tagged VACV. The experimental design incorporated a low multiplicity of infection, thereby enhancing detection of cellular proteins involved in cell-to-cell spread of VACV. The screen revealed 153 pro- and 149 anti-viral HFs that strongly influenced VACV replication. These HFs were investigated further by comparisons with transcriptional profiling data sets and HFs identified in RNAi screens of other viruses. In addition, functional and pathway analysis of the entire screen was carried out to highlight cellular mechanisms involved in VACV replication. This revealed, as anticipated, that many pro-viral HFs are involved in translation of mRNA and, unexpectedly, suggested that a range of proteins involved in cellular transcriptional processes and several DNA repair pathways possess anti-viral activity. Multiple components of the AMPK complex were found to act as pro-viral HFs, while several septins, a group of highly conserved GTP binding proteins with a role in sequestering intracellular bacteria, were identified as strong anti-viral VACV HFs. This screen has identified novel and previously unexplored roles for cellular factors in poxvirus replication. This advancement in our understanding of the VACV life cycle provides a reliable knowledge base for the improvement of poxvirus-based vaccine vectors and development of anti-viral theraputics.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine
Publisher:	Public Library of Science
ISSN:	1932-6203
Date of First Compliant Deposit:	8 November 2018
Date of Acceptance:	23 April 2014
Last Modified:	12 May 2023 00:35
URI:	https://orca.cardiff.ac.uk/id/eprint/110318

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