Evidence of a tick RNAi pathway by comparative genomics and reverse genetics screen of targets with known loss-of-function phenotypes in Drosophila

Kurscheid, Sebastian, Lew-Tabor, Ala E., Rodriguez Valle, Manuel, Bruyeres, Anthea G., Doogan, Vivienne J., Munderloh, Ulrike G., Guerrero, Felix D., Barrero, Roberto A. and Bellgard, Matthew I. (2009) Evidence of a tick RNAi pathway by comparative genomics and reverse genetics screen of targets with known loss-of-function phenotypes in Drosophila. BMC Molecular Biology, 10 .


Author Kurscheid, Sebastian
Lew-Tabor, Ala E.
Rodriguez Valle, Manuel
Bruyeres, Anthea G.
Doogan, Vivienne J.
Munderloh, Ulrike G.
Guerrero, Felix D.
Barrero, Roberto A.
Bellgard, Matthew I.
Title Evidence of a tick RNAi pathway by comparative genomics and reverse genetics screen of targets with known loss-of-function phenotypes in Drosophila
Formatted title Evidence of a tick RNAi pathway by comparative genomics and reverse genetics screen of targets with known loss-of-function phenotypes in Drosophila
Journal name BMC Molecular Biology   Check publisher's open access policy
ISSN 1471-2199
Publication date 2009
Sub-type Article (original research)
DOI 10.1186/1471-2199-10-26
Volume 10
Total pages 21
Place of publication London, United Kingdom
Publisher BioMed Central
Language eng
Formatted abstract Background: The Arthropods are a diverse group of organisms including Chelicerata (ticks, mites, spiders), Crustacea (crabs, shrimps), and Insecta (flies, mosquitoes, beetles, silkworm). The cattle tick, Rhipicephalus (Boophilus) microplus, is an economically significant ectoparasite of cattle affecting cattle industries world wide. With the availability of sequence reads from the first Chelicerate genome project (the Ixodes scapularis tick) and extensive R. microplus ESTs, we investigated evidence for putative RNAi proteins and studied RNA interference in tick cell cultures and adult female ticks targeting Drosophila homologues with known cell viability phenotype.
Results: We screened 13,643 R. microplus ESTs and I. scapularis genome reads to identify RNAi related proteins in ticks. Our analysis identified 31 RNAi proteins including a putative tick Dicer, RISC associated (Ago-2 and FMRp), RNA dependent RNA polymerase (EGO-1) and 23 homologues implicated in dsRNA uptake and processing. We selected 10 R. microplus ESTs with >80% similarity to D. melanogaster proteins associated with cell viability for RNAi functional screens in both BME26 R. microplus embryonic cells and female ticks in vivo. Only genes associated with proteasomes had an effect on cell viability in vitro. In vivo RNAi showed that 9 genes had significant effects either causing lethality or impairing egg laying.
Conclusion: We have identified key RNAi-related proteins in ticks and along with our loss-of-function studies support a functional RNAi pathway in R. microplus. Our preliminary studies indicate that tick RNAi pathways may differ from that of other Arthropods such as insects.
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ
Additional Notes Article # 26

Document type: Journal Article
Sub-type: Article (original research)
Collections: Queensland Alliance for Agriculture and Food Innovation
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Created: Thu, 13 Oct 2011, 10:27:12 EST by Dr Alicja Lew-tabor on behalf of Qld Alliance for Agriculture and Food Innovation