Identification of a novel leucine-rich repeat protein as a component of flagellar radial spoke in the Ascidian Ciona intestinalis

Mol Biol Cell. 2003 Feb;14(2):774-85. doi: 10.1091/mbc.02-06-0089.

Abstract

Axonemes are highly organized microtubule-based structures conserved in many eukaryotes. In an attempt to study axonemes by a proteomics approach, we selectively cloned cDNAs of axonemal proteins by immunoscreening the testis cDNA library from the ascidian Ciona intestinalis by using an antiserum against whole axonemes. We report here a 37-kDa protein of which cDNA occurred most frequently among total positive clones. This protein, named LRR37, belongs to the class of SDS22+ leucine-rich repeat (LRR) family. LRR37 is different from the LRR outer arm dynein light chain reported in Chlamydomonas and sea urchin flagella, and thus represents a novel axonemal LRR protein. Immunoelectron microscopy by using a polyclonal antibody against LRR37 showed that it is localized on the tip of the radial spoke, most likely on the spoke head. The LRR37 protein in fact seems to form a complex together with radial spoke protein 3 in a KI extract of the axonemes. These results suggest that LRR37 is a component of the radial spoke head and is involved in the interaction with other radial spoke components or proteins in the central pair projection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Western
  • Carrier Proteins / chemistry*
  • Carrier Proteins / genetics*
  • Chromatography, Gel
  • Ciona intestinalis
  • DNA, Complementary / metabolism
  • Electrophoresis, Gel, Two-Dimensional
  • Flagella / metabolism*
  • Gene Library
  • Immunohistochemistry
  • Leucine / chemistry*
  • Male
  • Microscopy, Fluorescence
  • Microscopy, Immunoelectron
  • Molecular Sequence Data
  • Protein Binding
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Testis / metabolism

Substances

  • Carrier Proteins
  • DNA, Complementary
  • Recombinant Fusion Proteins
  • Leucine