FEBS Letters
Volume 583, Issue 13 , Pages 2201-2207, 7 July 2009

Proteomic characterization of sperm radial spokes identifies a novel spoke protein with an ubiquitin domain

Edited by Dietmar J. Manstein

  • Yuhkoh Satouh

      Affiliations

    • Shimoda Marine Research Center, University of Tsukuba, Shimoda, Shizuoka 415-0025, Japan
    • Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan
    • ,
  • Kazuo Inaba

      Affiliations

    • Shimoda Marine Research Center, University of Tsukuba, Shimoda, Shizuoka 415-0025, Japan
    • Corresponding Author InformationCorresponding author. Fax: +81 558 22 0346.

Received 28 April 2009; received in revised form 2 June 2009; accepted 3 June 2009. published online 15 June 2009.

Abstract 

Radial spokes are T-shaped protein complexes important for the regulation of axonemal dyneins in eukaryotic cilia and flagella. Using a functional proteomics approach, we identified six spoke proteins in sperm flagella of the ascidian Ciona intestinalis. Many of the domain/motif structures in spoke proteins are commonly found in flagella of both Ciona sperm and Chlamydomonas, but interestingly they often distribute over non-orthologous protein components. A novel 116kDa protein named CMUB116 has both an ubiquitin domain and an IQ motif. It has orthologs in vertebrates, but not in Chlamydomonas. Furthermore, the results obtained by immunological analysis provide strong indication that CMUB116 is located at the stalk of radial spokes, where it is associated with MORN40.

Structured summary

MINT-7148244: CMUB116 (genbank_protein_gi:BAH59277) and MORN40 (genbank_protein_gi:BAH59284) colocalize (MI:0403) by cosedimentation (MI:0027)

MINT-7148179: Ci-RSP3 (uniprotkb:Q8T898) physically interacts (MI:0915) with tubulin alpha (uniprotkb:Q8MVT7), LRR37 (uniprotkb:Q8T896), CMUB116 (genbank_protein_gi:BAH59277), Ci-SRP4/6 (genbank_protein_gi:BAH59283), AK58 (genbank_protein_gi:BAM59278), tubulin beta (genbank_protein_gi:XP_002130315), NDK/DPY26 (genbank_protein_gi:BAH59279), MORN40 (genbank_protein_gi:BAH59284), ARM37 (genbank_protein_gi:BAH59280), NDK/DPY26 (genbank_protein_gi:NP_001161489), LC8 (genbank_protein_gi:BAH59282) and Ci-RSP9 (genbank_protein_gi:NP_001154962) by anti bait coimmunoprecipitation (MI:0006)

MINT-7148272: Ci-RSP3 (uniprotkb:Q8T898) and MORN40 (genbank_protein_gi:BAH59284) colocalize (MI:0403) by cosedimentation (MI:0027)

Abbreviations: MALDI-TOF/MS, matrix assisted laser desorption/ionization-time of flight mass spectrometry, RSP, radial spoke protein, LRR, leucine-rich repeat, NDK, nucleoside diphosphate kinase, MORN, membrane occupation and recognition nexus, ARM, armadillo repeat, LC, light chain

Keywords: Sperm flagella, Radial spoke, Proteomics, Ubiquitin, Nucleoside diphosphate kinase, Membrane occupation and recognition nexus, Ciona intestinalis

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PII: S0014-5793(09)00462-1

doi:10.1016/j.febslet.2009.06.016

FEBS Letters
Volume 583, Issue 13 , Pages 2201-2207, 7 July 2009

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