Loss of l(3)mbt leads to acquisition of the ping-pong cycle in Drosophila ovarian somatic cells

Genes Dev. 2016 Jul 15;30(14):1617-22. doi: 10.1101/gad.283929.116.

Abstract

In Drosophila germ cells, PIWI-interacting RNAs (piRNAs) are amplified through a PIWI slicer-dependent feed-forward loop termed the ping-pong cycle, yielding secondary piRNAs. However, the detailed mechanism remains poorly understood, largely because an ex vivo model system amenable to biochemical analyses has not been available. Here, we show that CRISPR-mediated loss of function of lethal (3) malignant brain tumor [l(3)mbt] leads to ectopic activation of the germ-specific ping-pong cycle in ovarian somatic cells. Perinuclear foci resembling nuage, the ping-pong center, appeared following l(3)mbt mutation. This activation of the ping-pong machinery in cultured cells will greatly facilitate elucidation of the mechanism underlying secondary piRNA biogenesis in Drosophila.

Keywords: CRISPR; Drosophila; PIWI; l(3)mbt; piRNA; ping-pong cycle.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Drosophila / genetics*
  • Drosophila Proteins / genetics
  • Drosophila Proteins / metabolism*
  • Female
  • Gene Silencing
  • Mutation
  • Ovary / cytology
  • Ovary / metabolism*
  • RNA, Small Interfering / biosynthesis
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism*

Substances

  • Drosophila Proteins
  • RNA, Small Interfering
  • l(3)mbt protein, Drosophila