Tudor staphylococcal nuclease (Tudor-SN) and Argonaute (Ago) are conserved components of the basic RNA interference (RNAi) machinery with a variety of functions including immune response and gene regulation. The RNAi machinery has been characterized in tick vectors of human and animal diseases but information is not available on the role of Tudor-SN in tick RNAi and other cellular processes. Our hypothesis is that tick Tudor-SN is part of the RNAi machinery and may be involved in innate immune response and other cellular processes. To address this hypothesis, Ixodes scapularis and I. ricinus ticks and/or cell lines were used to annotate and characterize the role of Tudor-SN in dsRNA-mediated RNAi, immune response to infection with the rickettsia Anaplasma phagocytophilum and the flaviviruses TBEV or LGTV and tick feeding. The results showed that Tudor-SN is conserved in ticks and involved in dsRNA-mediated RNAi and tick feeding but not in defense against infection with the examined viral and rickettsial pathogens. The effect of Tudor-SN gene knockdown on tick feeding could be due to down-regulation of genes that are required for protein processing and blood digestion through a mechanism that may involve selective degradation of dsRNAs enriched in G:U pairs that form as a result of adenosine-to-inosine RNA editing. These results demonstrated that Tudor-SN plays a role in tick RNAi pathway and feeding but no strong evidence for a role in innate immune responses to pathogen infection was found.

Department of Veterinary Pathobiology Center for Veterinary Health Sciences Oklahoma State University Stillwater Oklahoma United States of America

Institute of Parasitology Biology Centre Academy of Sciences of the Czech Republic Branišovská 31 37005 České Budějovice The Czech Republic

Institute of Virology Slovak Academy of Sciences Dúbravská cesta 9 84505 Bratislava Slovakia

Institute of Zoology Slovak Academy of Sciences Dúbravská cesta 9 84506 Bratislava Slovakia

SaBio Instituto de Investigación en Recursos Cinegéticos IREC CSIC UCLM JCCM Ronda de Toledo s n 13005 Ciudad Real Spain

SaBio Instituto de Investigación en Recursos Cinegéticos IREC CSIC UCLM JCCM Ronda de Toledo s n 13005 Ciudad Real Spain; Center for Infection and Immunity of Lille INSERM U1019 CNRS UMR 8204 Université Lille Nord de France Institut Pasteur de Lille Lille France

SaBio Instituto de Investigación en Recursos Cinegéticos IREC CSIC UCLM JCCM Ronda de Toledo s n 13005 Ciudad Real Spain; Department of Veterinary Pathobiology Center for Veterinary Health Sciences Oklahoma State University Stillwater Oklahoma United States of America

The Pirbright Institute Ash Road Pirbright Woking GU24 0NF United Kingdom

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