A Loss of Function Analysis of Host Factors Influencing Vaccinia virus Replication by RNA Interference
| Autor: | Hille Tekotte, Danielle K. Reynolds, Mike Tyers, Ralf Zimmer, Samantha J. Griffiths, Tali Pechenick Jowers, Jan Wildenhain, Manfred Auer, Orland Gonzalez, Philippa M. Beard, Peter Ghazal, Ismar R. Haga, Jürgen Haas |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Small interfering RNA
Transcription Genetic viruses lcsh:Medicine Pathology and Laboratory Medicine Virus Replication RNA interference Medicine and Health Sciences Vaccinia RNA Small Interfering lcsh:Science Genetics 0303 health sciences Multidisciplinary 030302 biochemistry & molecular biology virus diseases DNA virus Poxviruses General Medicine Cell biology Medical Microbiology Viral Pathogens Gene Knockdown Techniques Viruses Host-Pathogen Interactions RNA Interference Pathogens General Agricultural and Biological Sciences Research Article Signal Transduction DNA repair Vaccinia virus Biology Microbiology General Biochemistry Genetics and Molecular Biology 03 medical and health sciences Virology Humans Gene Microbial Pathogens 030304 developmental biology lcsh:R DNA replication Organisms Biology and Life Sciences Reproducibility of Results High-Throughput Screening Assays Viral replication Gene Expression Regulation lcsh:Q Genetic screen |
| Zdroj: | PLoS ONE PLoS ONE, Vol 9, Iss 6, p e98431 (2014) Beard, P M, Griffiths, S J, Gonzalez, O, Haga, I R, Pechenick-Jowers, T, Reynolds, D K, Wildenhain, J, Tekotte, H, Auer, M, Tyers, M, Ghazal, P, Zimmer, R & Haas, J 2014, ' A Loss of Function Analysis of Host Factors Influencing Vaccinia virus Replication by RNA Interference ', PLoS ONE, vol. 9, no. 6, e98431 . https://doi.org/10.1371/journal.pone.0098431 |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0098431 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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