Large Drosophila germline piRNA clusters are evolutionarily labile and dispensable for transposon regulation

Highlights

• Fly piRNA clusters are evolutionarily short lived and arise at unstable genomic loci

• Flies lacking the three major germline piRNA clusters are viable and fertile

• Despite loss of >40% of transposon-derived piRNAs, transposons are not activated

• Rather than by large clusters, regulation may be achieved in cis by dispersed elements

Summary

PIWI proteins and their guiding Piwi-interacting small RNAs (piRNAs) are crucial for fertility and transposon defense in the animal germline. In most species, the majority of piRNAs are produced from distinct large genomic loci, called piRNA clusters. It is assumed that germline-expressed piRNA clusters, particularly in Drosophila, act as principal regulators to control transposons dispersed across the genome. Here, using synteny analysis, we show that large clusters are evolutionarily labile, arise at loci characterized by recurrent chromosomal rearrangements, and are mostly species-specific across the Drosophila genus. By engineering chromosomal deletions in D. melanogaster, we demonstrate that the three largest germline clusters, which account for the accumulation of >40% of all transposon-targeting piRNAs in ovaries, are neither required for fertility nor for transposon regulation in trans. We provide further evidence that dispersed elements, rather than the regulatory action of large Drosophila germline clusters in trans, may be central for transposon defense.