Modulation of the transcriptional response of innate immune and RNAi genes upon exposure to dsRNA and LPS in silkmoth-derived Bm5 cells overexpressing BmToll9-1 receptor.
Injection or feeding of dsRNA is commonly used to induce specific gene silencing by RNAi in insects but very little research has been carried out to investigate non-specific effects on gene expression of dsRNA as pathogen-associated molecular pattern (PAMP). This study focuses on the potential role of the BmToll9-1 receptor to modulate the transcriptional response of innate immune and RNAi genes to dsRNA and lipopolysaccharide (LPS), which was used for comparison. To study this role, we took advantage of the silkmoth-derived Bm5 cell line, which does not express BmToll9-1 endogenously, and engineered a transformed cell line that permanently expresses BmToll9-1. Quantitative mRNA expression studies showed that BmToll9-1 can significantly alter the transcriptional response to dsRNA and LPS: (1) BmToll9-1 promotes the transcriptional response of Dicer2, encoding a key component of the RNAi machinery, and, to a lesser extent, that of transcription factors in the Jak-STAT and Toll pathways; and (2) BmToll9-1 represses the transcriptional induction of the IMD and Jak-STAT pathway genes, as well as the antimicrobial peptide (AMP) effector genes, by LPS. Thus, BmToll9-1 was identified as a modulator of innate immune and RNAi machinery gene expression that could be related to its preferential expression in the larval gut, the major barrier of pathogen entry. While BmToll9-1 was found to modulate RNAi-related gene expression, a reporter-based RNAi assay established no evidence for a direct interaction of BmToll9-1 with the intracellular RNAi machinery.
Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.
Jisheng
J
Liu
Insect Molecular Genetics and Biotechnology, Institute of Biosciences & Applications, National Centre for Scientific Research "Demokritos", Athens, Greece.
Anna
A
Kolliopoulou
Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium. Electronic address: guy.smagghe@ugent.be.
Guy
G
Smagghe
Insect Molecular Genetics and Biotechnology, Institute of Biosciences & Applications, National Centre for Scientific Research "Demokritos", Athens, Greece. Electronic address: swevers@bio.demokritos.gr.
Luc
L
Swevers
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D012831:Bombyx / Q000235:genetics* / Q000254:growth & development / Q000276:immunology*; D002460:Cell Line; D003001:Cloning, Molecular; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D007814:Larva / Q000235:genetics / Q000254:growth & development / Q000276:immunology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009928:Organ Specificity; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000494:pharmacology; D060888:Real-Time Polymerase Chain Reaction; D051217:Toll-Like Receptor 9 / Q000235:genetics* / Q000378:metabolism
D019476:Insect Proteins; D008070:Lipopolysaccharides; D012330:RNA, Double-Stranded; D051217:Toll-Like Receptor 9
AMP; Bm5; Bombyx mori; RNAi; Toll9-1; dsRNA