Structural organization of the intermediate and light chain complex of Chlamydomonas ciliary I1 dynein

Gang Fu, Chasity Scarbrough, Kangkang Song, Nhan Phan, Maureen Wirschell, Daniela Nicastro

Research output: Contribution to journalArticlepeer-review

Abstract

Axonemal I1 dynein (dynein f) is the largest inner dynein arm in cilia and a key regulator of ciliary beating. It consists of two dynein heavy chains, and an intermediate chain/light chain (ICLC) complex. However, the structural organization of the nine ICLC subunits remains largely unknown. Here, we used biochemical and genetic approaches, and cryo-electron tomography imaging in Chlamydomonas to dissect the molecular architecture of the I1 dynein ICLC complex. Using a strain expressing SNAP-tagged IC140, tomography revealed the location of the IC140 N-terminus at the proximal apex of the ICLC structure. Mass spectrometry of a tctex2b mutant showed that TCTEX2B dynein light chain is required for the stable assembly of TCTEX1 and inner dynein arm interacting proteins IC97 and FAP120. The structural defects observed in tctex2b located these 4 subunits in the center and bottom regions of the ICLC structure, which overlaps with the location of the IC138 regulatory subcomplex, which contains IC138, IC97, FAP120, and LC7b. These results reveal the three-dimensional organization of the native ICLC complex and indicate potential protein-protein interactions that are involved in the pathway by which I1 regulates ciliary motility.

Original languageEnglish (US)
Article numbere21646
JournalFASEB Journal
Volume35
Issue number6
DOIs
StatePublished - Jun 2021

Keywords

  • cryo-electron tomography
  • flagella
  • IC140
  • ICLC complex
  • TCTEX2B

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

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