Btk29A-mediated tyrosine phosphorylation of armadillo/β-catenin promotes ring canal growth in Drosophila oogenesis

PLoS One. 2015 Mar 24;10(3):e0121484. doi: 10.1371/journal.pone.0121484. eCollection 2015.

Abstract

Drosophila Btk29A is the ortholog of mammalian Btk, a Tec family nonreceptor tyrosine kinase whose deficit causes X-linked agammaglobulinemia in humans. The Btk29AficP mutation induces multiple abnormalities in oogenesis, including the growth arrest of ring canals, large intercellular bridges that allow the flow of cytoplasm carrying maternal products essential for embryonic development from the nurse cells to the oocyte during oogenesis. In this study, inactivation of Parcas, a negative regulator of Btk29A, was found to promote Btk29A accumulation on ring canals with a concomitant increase in the ring canal diameter, counteracting the Btk29AficP mutation. This mutation markedly reduced the accumulation of phosphotyrosine on ring canals and in the regions of cell-cell contact, where adhesion-supporting proteins such as DE-cadherin and β-catenin ortholog Armadillo (Arm) are located. Our previous in vitro and in vivo analyses revealed that Btk29A directly phosphorylates Arm, leading to its release from DE-cadherin. In the present experiments, immunohistological analysis revealed that phosphorylation at tyrosine 150 (Y150) and Y667 of Arm was diminished in Btk29AficP mutant ring canals. Overexpression of an Arm mutant with unphosphorylatable Y150 inhibited ring canal growth. Thus Btk29A-induced Y150 phosphorylation is necessary for the normal growth of ring canals. We suggest that the dissociation of tyrosine-phosphorylated Arm from DE-cadherin allows dynamic actin to reorganize, leading to ring canal expansion and cell shape changes during the course of oogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism
  • Animals
  • Armadillo Domain Proteins / chemistry*
  • Armadillo Domain Proteins / metabolism*
  • Drosophila Proteins / chemistry*
  • Drosophila Proteins / metabolism*
  • Drosophila melanogaster / cytology
  • Drosophila melanogaster / embryology*
  • Drosophila melanogaster / metabolism
  • Extracellular Space / metabolism*
  • Intracellular Space / metabolism
  • Oogenesis*
  • Phosphorylation
  • Protein Transport
  • Protein-Tyrosine Kinases / metabolism*
  • Transcription Factors / chemistry*
  • Transcription Factors / metabolism*
  • Tyrosine / metabolism*

Substances

  • ARM protein, Drosophila
  • Actins
  • Armadillo Domain Proteins
  • Drosophila Proteins
  • Transcription Factors
  • pcs protein, Drosophila
  • Tyrosine
  • Protein-Tyrosine Kinases
  • Btk29A protein, Drosophila

Grants and funding

This work was supported by Grants-in-Aid for Scientific Research (Nos. 23220007, 26113702 and 26114502) from MEXT to DY, a Life Science Grant from the Takeda Science Foundation to DY, a grant from The Ichiro Kanehara Foundation for the Promotion of Medical Sciences and Medical Care to NHK, a grant from The Scandinavia-Japan Sasakawa Foundation to NHK, a grant from the Tomizawa Jun-ichi & Keiko Fund of the Molecular Biology Society of Japan for Young Scientists to NHK, a grant from the Daiichi Sankyo Foundation of Life Science to NHK, and a Zoological MO Award from The Zoological Society of Japan to NHK. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.