The nexin link and B-tubule glutamylation maintain the alignment of outer doublets in the ciliary axoneme

Lea M. Alford, Daniel Stoddard, Jennifer H. Li, Emily L. Hunter, Douglas Tritschler, Raqual Bower, Daniela Nicastro, Mary E Porter, Winfield S. Sale

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

We developed quantitative assays to test the hypothesis that the N-DRC is required for integrity of the ciliary axoneme. We examined reactivated motility of demembranated drc cells, commonly termed “reactivated cell models.” ATP-induced reactivation of wild-type cells resulted in the forward swimming of ∼90% of cell models. ATP-induced reactivation failed in a subset of drc cell models, despite forward motility in live drc cells. Dark-field light microscopic observations of drc cell models revealed various degrees of axonemal splaying. In contrast, >98% of axonemes from wild-type reactivated cell models remained intact. The sup-pf4 and drc3 mutants, unlike other drc mutants, retain most of the N-DRC linker that interconnects outer doublet microtubules. Reactivated sup-pf4 and drc3 cell models displayed nearly wild-type levels of forward motility. Thus, the N-DRC linker is required for axonemal integrity. We also examined reactivated motility and axoneme integrity in mutants defective in tubulin polyglutamylation. ATP-induced reactivation resulted in forward swimming of >75% of tpg cell models. Analysis of double mutants defective in tubulin polyglutamylation and different regions of the N-DRC indicate B-tubule polyglutamylation and the distal lobe of the linker region are both important for axonemal integrity and normal N-DRC function.

Original languageEnglish (US)
Pages (from-to)331-340
Number of pages10
JournalCytoskeleton
Volume73
Issue number7
DOIs
StatePublished - Jun 1 2016

Bibliographical note

Publisher Copyright:
© 2016 Wiley Periodicals, Inc.

Keywords

  • DRC
  • N-DRC
  • axoneme
  • cilia
  • doublet microtubule
  • dynein
  • dynein regulatory complex
  • nexin

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