RNA-directed activation of cytoplasmic dynein-1 in reconstituted transport RNPs
Mark A McClintock,
Carly I Dix,
Christopher M Johnson,
Stephen H. McLaughlin,
Rory J Maizels,
Ha Thi Hoang,
Simon L Bullock
Posted 28 Feb 2018
bioRxiv DOI: 10.1101/273912 (published DOI: 10.7554/elife.36312)
Posted 28 Feb 2018
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 stabilises 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.
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