Transport and arrangement of the outer-dynein-arm docking complex in the flagella of Chlamydomonas mutants that lack outer dynein arms
UMass Chan Affiliations
Department of Cell BiologyDocument Type
Journal ArticlePublication Date
2001-03-29Keywords
AnimalsBiological Transport
Chlamydomonas reinhardtii
Cytoplasm
Dynein ATPase
Flagella
Fluorescent Antibody Technique
Microscopy, Immunoelectron
Mutation
Periodicity
Cell Biology
Metadata
Show full item recordAbstract
The outer dynein arms of Chlamydomonas flagella are attached to a precise site on the outer doublet microtubules and repeat at a regular interval of 24 nm. This binding is mediated by the outer dynein arm docking complex (ODA-DC), which is composed of three protein subunits. In this study, antibodies against the 83- and 62-kD subunits (DC83 and DC62) of the ODA-DC were used to analyze its state of association with outer arm components within the cytoplasm, and its localization in the axonemes of oda mutants. Immunoprecipitation indicates that DC83 and DC62 are preassembled within the cytoplasm, but that they are not associated with outer arm dynein. Both proteins are lost or greatly diminished in oda1 and oda3, mutants in the structural genes of DC62 and DC83, respectively, demonstrating that their association is necessary for their stable presence in the cytoplasm. Immunoelectron microscopy indicates that DC83 repeats at 24-nm intervals along the length of the doublet microtubules of oda6, which lacks outer arms; thus, outer arm periodicity may be determined by the ODA-DC. Flagellar regeneration and temporary dikaryon experiments indicate that the ODA-DC can be rapidly transported into the flagellum and assembled on the doublet microtubules independently of the outer arms and independently of flagellar growth. Unexpectedly, the intensity of ODA-DC labeling decreased toward the distal ends of axonemes of oda6 but not wild-type cells, suggesting that the outer arms reciprocally contribute to the assembly/stability of the ODA-DC.Source
Cell Motil Cytoskeleton. 2001 Apr;48(4):277-86. Link to article on publisher's siteDOI
10.1002/cm.1015Permanent Link to this Item
http://hdl.handle.net/20.500.14038/26523PubMed ID
11276076Related Resources
Link to Article in PubMedae974a485f413a2113503eed53cd6c53
10.1002/cm.1015