Continuum dynamics and statistical correction of compositional heterogeneity in multivalent IDP oligomers resolved by single-particle EM.

Autor: Mostofian B; Department of Biomedical Engineering, Oregon Health and Science University, Portland, OR 97239, USA., McFarland R; Department of Chemistry, Portland State University, Portland, OR 97201, USA., Estelle A; Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR 97331, USA., Howe J; Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR 97331, USA., Barbar E; Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR 97331, USA. Electronic address: elisar.barbar@oregonstate.edu., Reichow SL; Department of Chemistry, Portland State University, Portland, OR 97201, USA. Electronic address: reichow@pdx.edu., Zuckerman DM; Department of Biomedical Engineering, Oregon Health and Science University, Portland, OR 97239, USA. Electronic address: zuckermd@ohsu.edu.
Jazyk: angličtina
Zdroj: Journal of molecular biology [J Mol Biol] 2022 May 15; Vol. 434 (9), pp. 167520. Date of Electronic Publication: 2022 Mar 01.
DOI: 10.1016/j.jmb.2022.167520
Abstrakt: Multivalent intrinsically disordered protein (IDP) complexes are prevalent in biology and act in regulation of diverse processes, including transcription, signaling events, and the assembly and disassembly of complex macromolecular architectures. These systems pose significant challenges to structural investigation, due to continuum dynamics imparted by the IDP and compositional heterogeneity resulting from characteristic low-affinity interactions. Here, we developed a modular pipeline for automated single-particle electron microscopy (EM) distribution analysis of common but relatively understudied semi-ordered systems: 'beads-on-a-string' assemblies, composed of IDPs bound at multivalent sites to the ubiquitous ∼20 kDa cross-linking hub protein LC8. This approach quantifies conformational geometries and compositional heterogeneity on a single-particle basis, and statistically corrects spurious observations arising from random proximity of bound and unbound LC8. The statistical correction is generically applicable to oligomer characterization and not specific to our pipeline. Following validation, the approach was applied to the nuclear pore IDP Nup159 and the transcription factor ASCIZ. This analysis unveiled significant compositional and conformational diversity in both systems that could not be obtained from ensemble single particle EM class-averaging strategies, and new insights for exploring how these architectural properties might contribute to their physiological roles in supramolecular assembly and transcriptional regulation. We expect that this approach may be adopted to many other intrinsically disordered systems that have evaded traditional methods of structural characterization.
Competing Interests: Competing Interest Statement The authors have no competing interests.
(Copyright © 2022 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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