Endo-siRNAs depend on a new isoform of loquacious and target artificially introduced, high-copy sequences
Open Access
- 30 July 2009
- journal article
- research article
- Published by Springer Nature in The EMBO Journal
- Vol. 28 (19) , 2932-2944
- https://doi.org/10.1038/emboj.2009.220
Abstract
Colonization of genomes by a new selfish genetic element is detrimental to the host species and must lead to an efficient, repressive response. In vertebrates as well as in Drosophila , piRNAs repress transposons in the germ line, whereas endogenous siRNAs take on this role in somatic cells. We show that their biogenesis depends on a new isoform of the Drosophila TRBP homologue loquacious , which arises by alternative polyadenylation and is distinct from the one that functions during the biogenesis of miRNAs. For endo‐siRNAs and piRNAs, it is unclear how an efficient response can be initiated de novo . Our experiments establish that the endo‐siRNA pathway will target artificially introduced sequences without the need for a pre‐existing template in the genome. This response is also triggered in transiently transfected cells, thus genomic integration is not essential. Deep sequencing showed that corresponding endo‐siRNAs are generated throughout the sequence, but preferentially from transcribed regions. One strand of the dsRNA precursor can come from spliced mRNA, whereas the opposite strand derives from independent transcripts in antisense orientation.Keywords
This publication has 75 references indexed in Scilit:
- Specialized piRNA Pathways Act in Germline and Somatic Tissues of the Drosophila OvaryCell, 2009
- Collapse of Germline piRNAs in the Absence of Argonaute3 Reveals Somatic piRNAs in FliesCell, 2009
- Small RNAs as Guardians of the GenomeCell, 2009
- dsRNA Binding Properties of RDE-4 and TRBP Reflect Their Distinct Roles in RNAiJournal of Molecular Biology, 2008
- An Inside Job for siRNAsMolecular Cell, 2008
- Endogenous RNA Interference Provides a Somatic Defense against Drosophila TransposonsCurrent Biology, 2008
- Argonaute Loading Improves the 5′ Precision of Both MicroRNAs and Their miRNA∗ Strands in FliesCurrent Biology, 2008
- Sorting of Drosophila Small Silencing RNAsCell, 2007
- Drosophila microRNAs Are Sorted into Functionally Distinct Argonaute Complexes after Production by Dicer-1Cell, 2007
- Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT MethodMethods, 2001