Analysis of intraflagellar transport in C. elegans sensory cilia.
Cilia are assembled and maintained by intraflagellar transport (IFT), the motor-dependent, bidirectional movement of multiprotein complexes, called IFT particles, along the axoneme. The sensory cilia of Caenorhabditis elegans represent very useful objects for studying IFT because of the availability of in vivo time-lapse fluorescence microscopy assays of IFT and multiple ciliary mutants. In this system there are 60 sensory neurons, each having sensory cilia on the endings of their dendrites, and most components of the IFT machinery operating in these structures have been identified using forward and reverse genetic approaches. By analyzing the rate of IFT along cilia within living wild-type and mutant animals, two anterograde and one retrograde IFT motors were identified, the functional coordination of the two anterograde kinesin-2 motors was established and the transport properties of all the known IFT particle components have been characterized. The anterograde kinesin motors have been heterologously expressed and purified, and their biochemical properties have been characterized using MT gliding and single molecule motility assays. In this chapter, we summarize how the tools of genetics, cell biology, electron microscopy, and biochemistry are being used to dissect the composition and mechanism of action of IFT motors and IFT particles in C. elegans.
Hao
Limin
L
Molecular and Cell Biology, University of California, Davis, California 95616, USA.
Acar
Seyda
S
Evans
James
J
Ou
Guangshuo
G
Scholey
Jonathan M
JM
D000818:Animals; D054468:Axoneme / Q000378:metabolism / Q000648:ultrastructure; D001692:Biological Transport / Q000502:physiology*; D017173:Caenorhabditis elegans / Q000166:cytology* / Q000235:genetics* / Q000378:metabolism; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism; D002465:Cell Movement; D002923:Cilia / Q000378:metabolism / Q000648:ultrastructure; D005407:Flagella / Q000378:metabolism / Q000648:ultrastructure; D016547:Kinesins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D020409:Molecular Motor Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D009154:Mutation; D020033:Protein Isoforms / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D011984:Sensory Receptor Cells / Q000378:metabolism / Q000648:ultrastructure
D029742:Caenorhabditis elegans Proteins; D020409:Molecular Motor Proteins; D020033:Protein Isoforms; D011993:Recombinant Fusion Proteins; D016547:Kinesins