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E-GEOD-41780 - De novo piRNA cluster formation in the Drosophila germline triggered by transgenes containing a transcribed transposon fragment

Status
Released on 25 April 2013, last updated on 7 May 2013
Organism
Drosophila melanogaster
Samples (12)
Protocols (3)
Description
PIWI interacting RNAs (piRNAs) provide defense against transposable element (TE) expansion in the germline of metazoans. piRNAs are processed from the transcripts encoded by specialized heterochromatic clusters enriched in damaged copies of transposons. How these regions are recognized as a source of piRNAs is still elusive. The aim of this study is to determine how transgenes that contain a fragment of the LINE-like I transposon lead to an acquired TE resistance in Drosophila. We show that such transgenes, being inserted in unique euchromatic regions which normally do not produce small RNAs, become de novo bidirectional piRNA clusters that silence I-element activity in the germline. Strikingly, small RNAs of both polarities are generated from the entire transgene and flanking genomic sequences — not only from the transposon fragment. Chromatin immunoprecipitation analysis shows that in ovaries the trimethylated histone 3 lysine 9 (H3K9me3) mark associates with transgenes producing piRNAs. We show that transgene-derived hsp70 piRNAs stimulate in trans cleavage of cognate endogenous transcripts with subsequent processing of the non-homologous parts of these transcripts into piRNAs. The fractions of small RNAs (19-29 nt) from ovaries of wild type and 11 transgenic lines of Drosophila melanogaster were sequenced using Illumina HiSeq 2000.
Experiment type
RNA-seq of non coding RNA 
Contacts
Sergei Ryazansky <ryazansky@img.ras.ru>, Alla Kalmykova, Ivan Olovnikov, Sergey Shpiz
Citation
MINSEQE
Exp. designProtocolsVariablesProcessedSeq. reads
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