Abstract
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. 2009 Elsevier Inc. All rights reserved.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 235-266 |
| Number of pages | 32 |
| Journal | Methods in Cell Biology |
| Volume | 93 |
| DOIs | |
| State | Published - 2009 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Cell Biology
Cite this
Analysis of intraflagellar transport in C. elegans sensory cilia. / Hao, Limin; Acar, Seyda; Evans, James; Ou, Guangshuo; Scholey, Jonathan M.
In: Methods in Cell Biology, Vol. 93, 2009, p. 235-266.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Analysis of intraflagellar transport in C. elegans sensory cilia.
AU - Hao, Limin
AU - Acar, Seyda
AU - Evans, James
AU - Ou, Guangshuo
AU - Scholey, Jonathan M.
PY - 2009
Y1 - 2009
N2 - 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. 2009 Elsevier Inc. All rights reserved.
AB - 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. 2009 Elsevier Inc. All rights reserved.
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UR - http://www.scopus.com/inward/citedby.url?scp=77954676827&partnerID=8YFLogxK
U2 - 10.1016/S0091-679X(08)93013-2
DO - 10.1016/S0091-679X(08)93013-2
M3 - Article
C2 - 20409821
AN - SCOPUS:77954676827
VL - 93
SP - 235
EP - 266
JO - Methods in Cell Biology
JF - Methods in Cell Biology
SN - 0091-679X
ER -