mRNA transport and localization is a key aspect of posttranscriptional gene regulation. While the transport of many mRNAs is thought to occur through the recruitment of molecular motors, it has been a challenge to identify RNA-binding proteins (RBPs) that directly interact with motors by conventional assays. In order to identify RBPs and their specific domains that are responsible for recruiting a motor to transport granules, we have developed a single-molecule RNA mobility assay that enables quantifying the effect of a tethered RBP on the movement of an RNA. We demonstrate that tethering of RNAs to myosin or kinesin through their well-characterized interacting proteins results in quantitative differences in RNA mobility. This methodology provides a framework for identifying RBPs that mediate associations with motors.
CITATION STYLE
Bhaskar, V., Jia, M., & Chao, J. A. (2020). A single-molecule RNA mobility assay to identify proteins that link RNAs to molecular motors. In Methods in Molecular Biology (Vol. 2166, pp. 269–282). Humana Press Inc. https://doi.org/10.1007/978-1-0716-0712-1_16
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