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Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution
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Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution

Published on: January 16, 2017

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

Mark A McClintock1, Carly I Dix1, Christopher M Johnson2

  • 1Division of Cell Biology, MRC Laboratory of Molecular Biology, Cambridge, United Kingdom.

Elife
|June 27, 2018
PubMed

Insights

This study reveals that specific RNA sequences are essential for activating dynein motor protein-based mRNA transport. The RNA-binding protein Egalitarian (Egl) bridges RNA to Bicaudal-D (BicD), enabling motor complex recruitment.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Polarized mRNA transport is crucial for spatial protein synthesis but RNP composition and transport regulation remain unclear.
  • Understanding the molecular machinery behind mRNA transport is key to deciphering cellular organization.

Purpose of the Study:

  • To reconstitute and analyze the minimal components required for microtubule minus end-directed mRNA transport.
  • To elucidate the roles of adaptor proteins and RNA in regulating motor protein activity during transport.

Main Methods:

  • Reconstitution of mRNA transport using purified proteins and RNA.
  • Biochemical assays to study protein-protein and protein-RNA interactions.
  • Analysis of dynein motor activity in vitro.

Main Results:

  • A minimal ribonucleoprotein particle (RNP) for mRNA transport consists of Bicaudal-D (BicD), Egalitarian (Egl), and mRNA.
  • RNA is unexpectedly required for the robust activation of dynein motor motility.
  • A cis-acting RNA localization signal enhances Egl binding to BicD, facilitating dynein-dynactin recruitment.

Conclusions:

  • RNA acts as a crucial scaffold, inducing conformational changes in BicD via Egl to activate dynein-based transport.
  • This mechanism highlights RNA-mediated regulation of intracellular transport by controlling motor protein activity.
  • Cargo-induced scaffolding of adaptor protein complexes offers a generalizable model for transport regulation.
Keywords:
Bicaudal-DEgalitariancell biologydyneinin vitro reconstitutionmRNA transportmicrotubulenone

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