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HITS-CLIP analysis of Yb binding sites in Drosophila ovary cell line


ABSTRACT: piRNAs direct Piwi to repress transposons to maintain genome integrity in Drosophila ovarian somatic cells. piRNA maturation and association with Piwi occur at perinuclear Yb bodies, the centers of piRNA biogenesis. Here, we show that piRNA intermediates arising from the piRNA cluster flamenco (flam) concentrate into perinuclear foci adjacent to Yb bodies, termed Flam bodies. Although flam expression is not required for Yb body formation, Yb, the core component of Yb bodies, is required for Flam body formation. Abolishment of the RNA-binding activity of Yb disrupts both Yb bodies and Flam bodies. Loss of Zucchini, an endoribonuclease necessary for piRNA maturation, enlarges Flam bodies, which now superimpose with Yb bodies. Yb directly binds flam, but not neighboring protein-coding gene, transcripts. Thus, Yb integrates piRNA processing factors and piRNA intermediates into Yb bodies and Flam bodies, respectively, through direct binding to enhance piRNA biogenesis and formation of piRNA-inducing silencing complexes. HITS-CLIP was performed using OSC (Ovarian Somatic Cells). The antibody for Drosophila Yb, which was generated in this study, was used. Obtained CLIP tags were analyzed using illumina HiSeq200.

ORGANISM(S): Drosophila melanogaster

SUBMITTER: Haruhiko Siomi 

PROVIDER: E-GEOD-54875 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Yb integrates piRNA intermediates and processing factors into perinuclear bodies to enhance piRISC assembly.

Murota Yukiko Y   Ishizu Hirotsugu H   Nakagawa Shinichi S   Iwasaki Yuka W YW   Shibata Shinsuke S   Kamatani Miharu K MK   Saito Kuniaki K   Okano Hideyuki H   Siomi Haruhiko H   Siomi Mikiko C MC  

Cell reports 20140619 1


PIWI-interacting RNAs (piRNAs) direct Piwi to repress transposons and maintain genome integrity in Drosophila ovarian somatic cells. piRNA maturation and association with Piwi occur at perinuclear Yb bodies, the centers of piRNA biogenesis. Here, we show that piRNA intermediates arising from the piRNA cluster flamenco (flam) localize to perinuclear foci adjacent to Yb bodies, termed Flam bodies. RNAi-based screening of piRNA factors revealed that Flam body formation depends on Yb, the core compo  ...[more]

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