Kinesin-3 and dynein mediate microtubule-dependent co-transport of mRNPs and endosomes

Abstract

Long-distance transport of mRNAs is important in determining polarity in eukaryotes. Molecular motors shuttle large ribonucleoprotein complexes (mRNPs) containing RNA-binding proteins and associated factors along microtubules. However, precise mechanisms including the interplay of molecular motors and a potential connection to membrane trafficking remain elusive. Here, we solve the motor composition of transported mRNPs containing the RNA-binding protein Rrm4 of the pathogen Ustilago maydis. The underlying transport process determines the axis of polarity in infectious filaments. Plus end-directed Kin3, a Kinesin-3 type motor, mediates anterograde transport of mRNPs and is also present in transport units moving retrogradely. Split-dynein Dyn1/2 functions in retrograde movement of mRNPs. Plus end-directed conventional kinesin Kin1 is indirectly involved by transporting minus end-directed Dyn1/2 back to plus ends. Importantly, we additionally demonstrate that Rrm4-containing mRNPs co-localise with the t-SNARE Yup1 on shuttling endosomes and that functional endosomes are essential for mRNP movement. Either loss of Kin3 or removal of its lipid-binding pleckstrin homology domain abolish Rrm4-dependent movement without preventing co-localisation of Rrm4 and Yup1-positive endosomes. In summary, we uncovered the combination of motors required for mRNP shuttling along microtubules. Furthermore, intimately linked co-transport of endosomes and mRNPs suggests vesicle hitchhiking as novel mode of mRNP transport.