Distinct architecture and composition of mouse axonemal radial spoke head revealed by cryo-EM

Proc Natl Acad Sci U S A. 2021 Jan 26;118(4):e2021180118. doi: 10.1073/pnas.2021180118.

Abstract

The radial spoke (RS) heads of motile cilia and flagella contact projections of the central pair (CP) apparatus to coordinate motility, but the morphology is distinct for protozoa and metazoa. Here we show the murine RS head is compositionally distinct from that of Chlamydomonas Our reconstituted murine RS head core complex consists of Rsph1, Rsph3b, Rsph4a, and Rsph9, lacking Rsph6a and Rsph10b, whose orthologs exist in the protozoan RS head. We resolve its cryo-electron microscopy (cryo-EM) structure at 3.2-Å resolution. Our atomic model further reveals a twofold symmetric brake pad-shaped structure, in which Rsph4a and Rsph9 form a compact body extended laterally with two long arms of twisted Rsph1 β-sheets and potentially connected dorsally via Rsph3b to the RS stalk. Furthermore, our modeling suggests that the core complex contacts the periodic CP projections either rigidly through its tooth-shaped Rsph4a regions or elastically through both arms for optimized RS-CP interactions and mechanosignal transduction.

Keywords: cilia; cryo-EM; flagella; primary ciliary dyskinesia; radial spoke.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Surface
  • Axoneme / chemistry*
  • Axoneme / metabolism*
  • Chlamydomonas
  • Cilia
  • Cryoelectron Microscopy / methods*
  • Cytoskeletal Proteins / chemistry
  • DNA-Binding Proteins / chemistry
  • Epitopes
  • Flagella
  • HEK293 Cells
  • Humans
  • Mice
  • Models, Molecular
  • Mutation
  • Protein Conformation
  • Recombinant Proteins

Substances

  • Antigens, Surface
  • Cytoskeletal Proteins
  • DNA-Binding Proteins
  • Epitopes
  • NZ-1 testis-specific antigen, mouse
  • RSPH1 protein, human
  • RSPH4A protein, human
  • RSPH9 protein, human
  • Recombinant Proteins