The kinetic landscape of an RNA binding protein in cells View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2021-02-10

AUTHORS

Deepak Sharma, Leah L. Zagore, Matthew M. Brister, Xuan Ye, Carlos E. Crespo-Hernández, Donny D. Licatalosi, Eckhard Jankowsky

ABSTRACT

Gene expression in higher eukaryotic cells orchestrates interactions between thousands of RNA-binding proteins (RBPs) and tens of thousands of RNAs1. The kinetics by which RBPs bind to and dissociate from their RNA sites are critical for the coordination of cellular RNA-protein interactions2. However, these kinetic parameters have not been experimentally measured in cells. Here we show that time-resolved RNA-protein cross-linking with a pulsed femtosecond ultraviolet laser, followed by immunoprecipitation and high-throughput sequencing, allows the determination of binding and dissociation kinetics of the RBP DAZL for thousands of individual RNA-binding sites in cells. This kinetic cross-linking and immunoprecipitation (KIN-CLIP) approach reveals that DAZL resides at individual binding sites for time periods of only seconds or shorter, whereas the binding sites remain DAZL-free for markedly longer. The data also indicate that DAZL binds to many RNAs in clusters of multiple proximal sites. The effect of DAZL on mRNA levels and ribosome association correlates with the cumulative probability of DAZL binding in these clusters. Integrating kinetic data with mRNA features quantitatively connects DAZL-RNA binding to DAZL function. Our results show how kinetic parameters for RNA-protein interactions can be measured in cells, and how these data link RBP-RNA binding to the cellular function of RBPs. More... »

PAGES

152-156

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/s41586-021-03222-x

DOI

http://dx.doi.org/10.1038/s41586-021-03222-x

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1135290988

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/33568810


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