Structural basis for high-affinity actin binding revealed by a β-III-spectrin SCA5 missense mutation

Adam W. Avery, Michael E. Fealey, Fengbin Wang, Albina Orlova, Andrew R. Thompson, David D. Thomas, Thomas S. Hays, Edward H. Egelman

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17 Scopus citations

Abstract

Spinocerebellar ataxia type 5 (SCA5) is a neurodegenerative disease caused by mutations in the cytoskeletal protein β-III-spectrin. Previously, a SCA5 mutation resulting in a leucine-to-proline substitution (L253P) in the actin-binding domain (ABD) was shown to cause a 1000-fold increase in actin-binding affinity. However, the structural basis for this increase is unknown. Here, we report a 6.9 Å cryo-EM structure of F-actin complexed with the L253P ABD. This structure, along with co-sedimentation and pulsed-EPR measurements, demonstrates that high-affinity binding caused by the CH2-localized mutation is due to opening of the two CH domains. This enables CH1 to bind actin aided by an unstructured N-terminal region that becomes α-helical upon binding. This helix is required for association with actin as truncation eliminates binding. Collectively, these results shed light on the mechanism by which β-III-spectrin, and likely similar actin-binding proteins, interact with actin, and how this mechanism can be perturbed to cause disease.

Original languageEnglish (US)
Article number1350
JournalNature communications
Volume8
Issue number1
DOIs
StatePublished - Dec 1 2017

Bibliographical note

Funding Information:
This work was partially supported by grants from NIH to T.S.H. (GM44757), D.D.T. (AG32961), and E.H.E. (GM81303), and from NSF (MCB-1616854 to D.D.T.). A.W.A. was supported by an NIH grant (GM44757) to T.S.H. and an NIH training grant to D.D.T. (T32 AR007612). M.E.F. was supported by an NIH training grant to D.D.T. (T32 AR007612). The cryo-EM work was conducted at the Molecular Electron Microscopy Core facility at the University of Virginia, which is supported by the School of Medicine and built with NIH grant G20-RR31199. The Titan Krios and Falcon II direct electron detector within the Core were purchased with NIH SIG S10-RR025067 and S10-OD018149, respectively.

Publisher Copyright:
© 2017 The Author(s).

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