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

Miki Imanishi, Nicholas F. Endres, Arne Gennerich, Ronald D. Vale

Research output: Contribution to journalArticle

63 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)931-937
Number of pages7
JournalJournal of Cell Biology
Volume174
Issue number7
DOIs
StatePublished - Sep 25 2006
Externally publishedYes

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Caenorhabditis elegans
Microtubules
Adenosine Triphosphatases
Optical Tweezers
Kinesin
Mutation
Cilia
Point Mutation
Fluorescence
Alleles
Maintenance

ASJC Scopus subject areas

  • Cell Biology

Cite this

Autoinhibition regulates the motility of the C. elegans intraflagellar transport motor OSM-3. / Imanishi, Miki; Endres, Nicholas F.; Gennerich, Arne; Vale, Ronald D.

In: Journal of Cell Biology, Vol. 174, No. 7, 25.09.2006, p. 931-937.

Research output: Contribution to journalArticle

Imanishi, Miki ; Endres, Nicholas F. ; Gennerich, Arne ; Vale, Ronald D. / Autoinhibition regulates the motility of the C. elegans intraflagellar transport motor OSM-3. In: Journal of Cell Biology. 2006 ; Vol. 174, No. 7. pp. 931-937.
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