Building blocks of the nexin-dynein regulatory complex in chlamydomonas flagella

Jianfeng Lin, Douglas Tritschler, Kangkang Song, Cynthia F. Barber, Jennifer S. Cobb, Mary E. Porter, Daniela Nicastro

Research output: Contribution to journalArticle

58 Scopus citations

Abstract

The directional flow generated by motile cilia and flagella is critical for many processes, including human development and organ function. Normal beating requires the control and coordination of thousands of dynein motors, and the nexin-dynein regulatory complex (N-DRC) has been identified as an important regulatory node for orchestrating dynein activity. The nexin link appears to be critical for the transformation of dynein-driven, linear microtubule sliding to flagellar bending, yet the molecular composition and mechanism of the N-DRC remain largely unknown. Here, we used proteomics with special attention to protein phosphorylation to analyze the composition of the N-DRC and to determine which subunits may be important for signal transduction. Two-dimensional electrophoresis and MALDI-TOF mass spectrometry of WT and mutant flagellar axonemes from Chlamydomonas identified 12 N-DRC-associated proteins, including all seven previously observed N-DRC components. Sequence and PCR analyses identified the mutation responsible for the phenotype of the sup-pf-4 strain, and biochemical comparison with a radial spoke mutant revealed two components that may link the N-DRC and the radial spokes. Phosphoproteomics revealed eight proteins with phosphorylated isoforms for which the isoform patterns changed with the genotype as well as two components that may play pivotal roles in N-DRC function through their phosphorylation status. These data were assembled into a model of the N-DRC that explains aspects of its regulatory function.

Original languageEnglish (US)
Pages (from-to)29175-29191
Number of pages17
JournalJournal of Biological Chemistry
Volume286
Issue number33
DOIs
StatePublished - Aug 19 2011

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Lin, J., Tritschler, D., Song, K., Barber, C. F., Cobb, J. S., Porter, M. E., & Nicastro, D. (2011). Building blocks of the nexin-dynein regulatory complex in chlamydomonas flagella. Journal of Biological Chemistry, 286(33), 29175-29191. https://doi.org/10.1074/jbc.M111.241760