Systems NMR: single-sample quantification of RNA, proteins and metabolites for biomolecular network analysis
Abstract
Cellular behavior is controlled by the interplay of diverse biomolecules. Most experimental methods, however, can only monitor a single molecule class or reaction type at a time. We developed an in vitro nuclear magnetic resonance spectroscopy (NMR) approach, which permitted dynamic quantification of an entire ‘heterotypic’ network—simultaneously monitoring three distinct molecule classes (metabolites, proteins and RNA) and all elementary reaction types (bimolecular interactions, catalysis, unimolecular changes). Focusing on an eight-reaction co-transcriptional RNA folding network, in a single sample we recorded over 35 time points with over 170 observables each, and accurately determined five core reaction constants in multiplex. This reconstruction revealed unexpected cross-talk between the different reactions. We further observed dynamic phase-separation in a system of five distinct RNA-binding domains in the course of the RNA transcription reaction. Our Systems NMR approach provides a deeper understanding of biological network dynamics by combining the dynamic resolution of biochemical assays and the multiplexing ability of ‘omics’. Show more
Permanent link
https://doi.org/10.3929/ethz-b-000357247Publication status
publishedExternal links
Journal / series
Nature MethodsVolume
Pages / Article No.
Publisher
NatureSubject
Analytical biochemistry; Biochemical reaction networks; NMR spectroscopy; RNA; Systems biologyOrganisational unit
03791 - Iber, Dagmar / Iber, Dagmar
03927 - Picotti, Paola / Picotti, Paola
03591 - Allain, Frédéric / Allain, Frédéric
Notes
It was possible to publish this article open access thanks to a Swiss National Licence with the publisher.More
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