Role of ADMA-histones in dual-strand piRNA source loci recognition by Rhino

In Drosophila germ cells, piRNAs arise from dual-strand piRNA clusters, which are marked by a repressive histone mark H3K9me3, but are transcribed from internal sites in a manner dependent on the HP1 homolog Rhino. Rhino binds to H3K9me3 on these clusters, yet the mechanism controlling its binding to clusters remains unclear. Here, we used cultured ovarian somatic cells (OSCs), which lack endogenous Rhino and its stabilizer Kipferl, whose absence renders the dual-strand clusters inert, and found that exogenous Rhino tends to bind to the ends of dual-strand clusters with asymmetric dimethylarginine histones (ADMA-histones). Depletion of the arginine methyltransferases responsible for ADMA modifications, such as DART4, affected the genomic localization of Rhino in both OSCs and ovaries. We also identified primitive, cluster-like genomic regions, termed DART4 piSL, where Rhino propagates in an DART4-dependent but unstable manner. Our study proposes that ADMA-histones play a crucial role in the initial genome loading of Rhino and may establish the potential sites of its propagation, which are subsequently stabilized to support piRNA production.