Autoinhibition regulates the motility of the C. elegans intraflagellar transport motor OSM-3

J Cell Biol. 2006 Sep 25;174(7):931-7. doi: 10.1083/jcb.200605179.

Abstract

OSM-3 is a Kinesin-2 family member from Caenorhabditis elegans that is involved in intraflagellar transport (IFT), a process essential for the construction and maintenance of sensory cilia. In this study, using a single-molecule fluorescence assay, we show that bacterially expressed OSM-3 in solution does not move processively (multiple steps along a microtubule without dissociation) and displays low microtubule-stimulated adenosine triphosphatase (ATPase) activity. However, a point mutation (G444E) in a predicted hinge region of OSM-3's coiled-coil stalk as well as a deletion of that hinge activate ATPase activity and induce robust processive movement. These hinge mutations also cause a conformational change in OSM-3, causing it to adopt a more extended conformation. The motility of wild-type OSM-3 also can be activated by attaching the motor to beads in an optical trap, a situation that may mimic attachment to IFT cargo. Our results suggest that OSM-3 motility is repressed by an intramolecular interaction that involves folding about a central hinge and that IFT cargo binding relieves this autoinhibition in vivo. Interestingly, the G444E allele in C. elegans produces similar ciliary defects to an osm-3-null mutation, suggesting that autoinhibition is important for OSM-3's biological function.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / metabolism
  • Animals
  • Biological Transport
  • Caenorhabditis elegans / physiology*
  • Caenorhabditis elegans Proteins / genetics
  • Caenorhabditis elegans Proteins / metabolism
  • Caenorhabditis elegans Proteins / physiology*
  • DNA, Complementary / biosynthesis
  • Down-Regulation*
  • Flagella / physiology*
  • Kinesins / genetics
  • Kinesins / metabolism
  • Kinesins / physiology*
  • Microscopy, Fluorescence
  • Models, Biological
  • Molecular Motor Proteins / physiology*
  • Protein Folding
  • Recombinant Proteins / genetics

Substances

  • Caenorhabditis elegans Proteins
  • DNA, Complementary
  • Molecular Motor Proteins
  • OSM-3 protein, C elegans
  • Recombinant Proteins
  • Adenosine Triphosphatases
  • Kinesins