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PMID: 19752022 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Regulation of dynein-driven microtubule sliding by the axonemal protein kinase CK1 in Chlamydomonas flagella.

The Journal of cell biology ·Vol. 186 ·No. 6 ·2009-09-21 ·Pages 817-24

Gokhale A, Wirschell M, Sale WS

Abstract

Experimental analysis of isolated ciliary/flagellar axonemes has implicated the protein kinase casein kinase I (CK1) in regulation of dynein. To test this hypothesis, we developed a novel in vitro reconstitution approach using purified recombinant Chlamydomonas reinhardtii CK1, together with CK1-depleted axonemes from the paralyzed flagellar mutant pf17, which is defective in radial spokes and impaired in dynein-driven microtubule sliding. The CK1 inhibitors (DRB and CK1-7) and solubilization of CK1 restored microtubule sliding in pf17 axonemes, which is consistent with an inhibitory role for CK1. The phosphatase inhibitor microcystin-LR blocked rescue of microtubule sliding, indicating that the axonemal phosphatases, required for rescue, were retained in the CK1-depleted axonemes. Reconstitution of depleted axonemes with purified, recombinant CK1 restored inhibition of microtubule sliding in a DRB- and CK1-7-sensitive manner. In contrast, a purified "kinase-dead" CK1 failed to restore inhibition. These results firmly establish that an axonemal CK1 regulates dynein activity and flagellar motility.

MeSH Terms
Animals Axoneme/drug effects,enzymology Casein Kinase I/antagonists & inhibitors,genetics,metabolism Cell Movement/drug effects Chlamydomonas reinhardtii/drug effects,enzymology,genetics Dichlororibofuranosylbenzimidazole/pharmacology Dyneins/metabolism Flagella/drug effects,enzymology Isoquinolines/pharmacology Marine Toxins Microcystins/pharmacology Mutation Phosphoprotein Phosphatases/antagonists & inhibitors Protein Kinase Inhibitors/pharmacology Recombinant Proteins/metabolism
Chemicals
Isoquinolines Marine Toxins Microcystins Protein Kinase Inhibitors Recombinant Proteins N-(2-aminoethyl)-5-chloroisoquinoline-8-sulfonamide Dichlororibofuranosylbenzimidazole Casein Kinase I Phosphoprotein Phosphatases Dyneins cyanoginosin LR
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Gokhale Avanti
Department of Cell Biology, Emory University School of Medicine, Atlanta, GA 30322, USA.
Wirschell Maureen
Sale Winfield S
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
1540-8140
DOI
10.1083/jcb.200906168
Published
2009-09-21
Epub
2009-00-14
Pages
817-24
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2753152
Subset
IM
Grants
NIGMS NIH HHS · R01 GM051173 · United States
NIGMS NIH HHS · GM075446 · United States
NEI NIH HHS · EY070911 · United States
NIGMS NIH HHS · R37 GM051173 · United States
NIGMS NIH HHS · GM051173 · United States
NIGMS NIH HHS · F32 GM075446 · United States
External Links
PubMed source DOI: 10.1083/jcb.200906168 PMC Full Text View journal details
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