期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 116, 期 46, 页码 23152-23162出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1910960116
关键词
cilia; flagella; cryo-electron tomography; N-DRC; axoneme
资金
- Cancer Prevention and Research Institute of Texas (CPRIT) [RP170644]
- National Science Foundation (NSF) [9871237, NSF-DBI-0215759]
- NSF Living Stock Collections for Biological Research Program [0951671, 00017383]
- National Institutes of Health [R01GM083122, R01GM055667]
- CPRIT Grant [RR140082]
- Direct For Biological Sciences
- Div Of Biological Infrastructure [0951671] Funding Source: National Science Foundation
The nexin-dynein regulatory complex (N-DRC) in motile cilia and flagella functions as a linker between neighboring doublet microtubules, acts to stabilize the axonemal core structure, and serves as a central hub for the regulation of ciliary motility. Although the N-DRC has been studied extensively using genetic, biochemical, and structural approaches, the precise arrangement of the 11 (or more) N-DRC subunits remains unknown. Here, using cryo-electron tomography, we have compared the structure of Chlamydomonas wild-type flagella to that of strains with specific DRC subunit deletions or rescued strains with tagged DRC subunits. Our results show that DRC7 is a central linker subunit that helps connect the N-DRC to the outer dynein arms. DRC11 is required for the assembly of DRC8, and DRC8/11 form a subcomplex in the proximal lobe of the linker domain that is required to form stable contacts to the neighboring B-tubule. Gold labeling of tagged subunits determines the precise locations of the previously ambiguous N terminus of DRC4 and C terminus of DRC5. DRC4 is now shown to contribute to the core scaffold of the N-DRC. Our results reveal the overall architecture of N-DRC, with the 3 subunits DRC1/2/4 forming a core complex that serves as the scaffold for the assembly of the functional subunits, namely DRC3/5-8/11. These findings shed light on N-DRC assembly and its role in regulating flagellar beating.
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