Identification of PNG kinase substrates uncovers interactions with the translational repressor TRAL in the oocyte-to-embryo transition

Elife. 2018 Feb 26:7:e33150. doi: 10.7554/eLife.33150.

Abstract

The Drosophila Pan Gu (PNG) kinase complex regulates hundreds of maternal mRNAs that become translationally repressed or activated as the oocyte transitions to an embryo. In a previous paper (Hara et al., 2017), we demonstrated PNG activity is under tight developmental control and restricted to this transition. Here, examination of PNG specificity showed it to be a Thr-kinase yet lacking a clear phosphorylation site consensus sequence. An unbiased biochemical screen for PNG substrates identified the conserved translational repressor Trailer Hitch (TRAL). Phosphomimetic mutation of the PNG phospho-sites in TRAL reduced its ability to inhibit translation in vitro. In vivo, mutation of tral dominantly suppressed png mutants and restored Cyclin B protein levels. The repressor Pumilio (PUM) has the same relationship with PNG, and we also show that PUM is a PNG substrate. Furthermore, PNG can phosphorylate BICC and ME31B, repressors that bind TRAL in cytoplasmic RNPs. Therefore, PNG likely promotes translation at the oocyte-to-embryo transition by phosphorylating and inactivating translational repressors.

Keywords: BICC; D. melanogaster; Drosophila; ME31B; Pumilio; cytoplasmic RNPs; developmental biology; maternal mRNA; stem cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Drosophila / embryology*
  • Drosophila / enzymology*
  • Drosophila Proteins / metabolism*
  • Gene Expression Regulation, Developmental
  • Phosphorylation
  • Protein Interaction Mapping
  • Protein Processing, Post-Translational*
  • Protein Serine-Threonine Kinases / metabolism*
  • Ribonucleoproteins / metabolism*

Substances

  • Drosophila Proteins
  • Ribonucleoproteins
  • TRAL protein, Drosophila
  • png protein, Drosophila
  • Protein Serine-Threonine Kinases