RNA-directed activation of cytoplasmic dynein-1 in reconstituted transport RNPs

Elife. 2018 Jun 26:7:e36312. doi: 10.7554/eLife.36312.

Abstract

Polarised mRNA transport is a prevalent mechanism for spatial control of protein synthesis. However, the composition of transported ribonucleoprotein particles (RNPs) and the regulation of their movement are poorly understood. We have reconstituted microtubule minus end-directed transport of mRNAs using purified components. A Bicaudal-D (BicD) adaptor protein and the RNA-binding protein Egalitarian (Egl) are sufficient for long-distance mRNA transport by the dynein motor and its accessory complex dynactin, thus defining a minimal transport-competent RNP. Unexpectedly, the RNA is required for robust activation of dynein motility. We show that a cis-acting RNA localisation signal promotes the interaction of Egl with BicD, which licenses the latter protein to recruit dynein and dynactin. Our data support a model for BicD activation based on RNA-induced occupancy of two Egl-binding sites on the BicD dimer. Scaffolding of adaptor protein assemblies by cargoes is an attractive mechanism for regulating intracellular transport.

Keywords: Bicaudal-D; Egalitarian; cell biology; dynein; in vitro reconstitution; mRNA transport; microtubule; none.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • Cytoplasmic Dyneins / chemistry
  • Cytoplasmic Dyneins / genetics
  • Drosophila Proteins / chemistry
  • Drosophila Proteins / genetics*
  • Drosophila melanogaster / genetics
  • Dynactin Complex / chemistry
  • Dynactin Complex / genetics*
  • Dyneins / chemistry
  • Dyneins / genetics*
  • Protein Binding / genetics
  • Protein Multimerization
  • Protein Transport / genetics
  • RNA Transport / genetics
  • RNA, Messenger / genetics
  • RNA-Binding Proteins / chemistry
  • RNA-Binding Proteins / genetics
  • Ribonucleoproteins / genetics

Substances

  • BicD protein, Drosophila
  • Drosophila Proteins
  • Dynactin Complex
  • RNA, Messenger
  • RNA-Binding Proteins
  • Ribonucleoproteins
  • egl protein, Drosophila
  • Cytoplasmic Dyneins
  • Dyneins