PMID- 30422992 OWN - NLM STAT- MEDLINE DCOM- 20190215 LR - 20240404 IS - 1553-7374 (Electronic) IS - 1553-7366 (Print) IS - 1553-7366 (Linking) VI - 14 IP - 11 DP - 2018 Nov TI - Whole genome screen reveals a novel relationship between Wolbachia levels and Drosophila host translation. PG - e1007445 LID - 10.1371/journal.ppat.1007445 [doi] LID - e1007445 AB - Wolbachia is an intracellular bacterium that infects a remarkable range of insect hosts. Insects such as mosquitos act as vectors for many devastating human viruses such as Dengue, West Nile, and Zika. Remarkably, Wolbachia infection provides insect hosts with resistance to many arboviruses thereby rendering the insects ineffective as vectors. To utilize Wolbachia effectively as a tool against vector-borne viruses a better understanding of the host-Wolbachia relationship is needed. To investigate Wolbachia-insect interactions we used the Wolbachia/Drosophila model that provides a genetically tractable system for studying host-pathogen interactions. We coupled genome-wide RNAi screening with a novel high-throughput fluorescence in situ hybridization (FISH) assay to detect changes in Wolbachia levels in a Wolbachia-infected Drosophila cell line JW18. 1117 genes altered Wolbachia levels when knocked down by RNAi of which 329 genes increased and 788 genes decreased the level of Wolbachia. Validation of hits included in depth secondary screening using in vitro RNAi, Drosophila mutants, and Wolbachia-detection by DNA qPCR. A diverse set of host gene networks was identified to regulate Wolbachia levels and unexpectedly revealed that perturbations of host translation components such as the ribosome and translation initiation factors results in increased Wolbachia levels both in vitro using RNAi and in vivo using mutants and a chemical-based translation inhibition assay. This work provides evidence for Wolbachia-host translation interaction and strengthens our general understanding of the Wolbachia-host intracellular relationship. FAU - Grobler, Yolande AU - Grobler Y AD - Department of Cell Biology, Howard Hughes Medical Institute and Kimmel Center for Biology and Medicine at the Skirball Institute, New York University School of Medicine, New York, NY, United States of America. FAU - Yun, Chi Y AU - Yun CY AD - High Throughput Biology Core, Skirball Institute at New York University Langone Medical Center, New York, NY, United States of America. FAU - Kahler, David J AU - Kahler DJ AUID- ORCID: 0000-0002-7144-2215 AD - High Throughput Biology Core, Skirball Institute at New York University Langone Medical Center, New York, NY, United States of America. FAU - Bergman, Casey M AU - Bergman CM AUID- ORCID: 0000-0002-5462-9854 AD - Department of Genetics and Institute of Bioinformatics, University of Georgia, Athens, GA, United States of America. FAU - Lee, Hangnoh AU - Lee H AUID- ORCID: 0000-0003-0442-8330 AD - Section of Developmental Genomics, Laboratory of Cellular and Developmental Biology, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD, United States of America. FAU - Oliver, Brian AU - Oliver B AD - Section of Developmental Genomics, Laboratory of Cellular and Developmental Biology, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD, United States of America. FAU - Lehmann, Ruth AU - Lehmann R AUID- ORCID: 0000-0002-8454-5651 AD - Department of Cell Biology, Howard Hughes Medical Institute and Kimmel Center for Biology and Medicine at the Skirball Institute, New York University School of Medicine, New York, NY, United States of America. LA - eng GR - P30 CA016087/CA/NCI NIH HHS/United States GR - P40 OD018537/OD/NIH HHS/United States GR - HHMI/Howard Hughes Medical Institute/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20181113 PL - United States TA - PLoS Pathog JT - PLoS pathogens JID - 101238921 SB - IM MH - Animals MH - Culicidae MH - Drosophila/genetics/microbiology MH - Drosophila melanogaster/*genetics/microbiology MH - Genome MH - Host Microbial Interactions/*genetics MH - Host-Pathogen Interactions/genetics MH - Humans MH - In Situ Hybridization, Fluorescence/methods MH - Mosquito Vectors MH - RNA Interference MH - Symbiosis MH - Viruses/genetics MH - Whole Genome Sequencing/methods MH - Wolbachia/*genetics PMC - PMC6258568 COIS- The authors have declared that no competing interests exist. EDAT- 2018/11/14 06:00 MHDA- 2019/02/16 06:00 PMCR- 2018/11/13 CRDT- 2018/11/14 06:00 PHST- 2018/07/14 00:00 [received] PHST- 2018/10/30 00:00 [accepted] PHST- 2018/11/27 00:00 [revised] PHST- 2018/11/14 06:00 [pubmed] PHST- 2019/02/16 06:00 [medline] PHST- 2018/11/14 06:00 [entrez] PHST- 2018/11/13 00:00 [pmc-release] AID - PPATHOGENS-D-18-01403 [pii] AID - 10.1371/journal.ppat.1007445 [doi] PST - epublish SO - PLoS Pathog. 2018 Nov 13;14(11):e1007445. doi: 10.1371/journal.ppat.1007445. eCollection 2018 Nov.