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A single miRNA drives hemimetabolan metamophosis to a right end

AutorBellés, Xavier ; Lozano Fernández, Jesús ; Montañez, Raúl
Fecha de publicaciónjul-2013
CitaciónInsect Hormones (19th Ecdysone) International Workshop (2013)
ResumenPrevious studies carried out in our laboratory had shown that RNAi depletion of dicer-1, a key enzyme involved in miRNA maturation, inhibited metamorphosis in the cockroach Blattella germanica. Dicer-1 suppression in the last (6th) instar nymph triggered the depletion of miRNA expression, and the nymphs molted to a supernumerary 7th nymphal instar instead to molt to adults. In order to identify which miRNAs are involved in regulating metamorphosis, we focused our attention on Krüppel-h1 (Kr-h1), a JH-dependent transcription factor that represses metamorphosis. Indeed, a decrease of Kr-h1 expression in the pre-metamorphic instar is crucial for metamorphosis progression. The decrease is abrupt, and this abruptness is hardly explained only by mechanisms controlling Kr-h1 transcription, and might be driven by miRNAs. Using again B. germanica as model, we observed that RNAi of Kr-h1 carried out in dicer-1-depleted specimens rescued normal metamorphosis. This pointed to miRNAs affecting Kr-h1 as responsible of the metamorphosis inhibition triggered by dicer-1 depletion. We also observed that the 3’UTR of Kr-h1 mRNA contains predicted sites for miR-2 family miRNAs, and that the predicted sites are functional, according to the results of luciferase reporter assays. Moreover, depletion of miR-2a impaired the decrease of Kr-h1 expression leading to metamorphosis inhibition, and administration of miR-2a mimic rescued the normal metamorphosis phenotype in dicer-1-depleted insects. The whole data suggests that miR-2a plays the key role of radically removing Kr-h1 mRNA in the premetamorphic stage, as a prerequisite for the activation of the transcription factors repressed by Kr-h1 that are the ultimate inducers of metamorphosis.
DescripciónTrabajo presentado en el Insect Hormones (19th Ecdysone) International Workshop, celebrado en Minneapolis del 21 al 26 de julio de 2013.
URIhttp://hdl.handle.net/10261/153624
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