Flagellar Radial Spoke Protein 2 Is a Calmodulin Binding Protein Required for Motility in <i>Chlamydomonas reinhardtii</i>
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- Pinfen Yang
- Department of Biology, Marquette University, Milwaukee, Wisconsin 53233
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- Chun Yang
- Department of Biology, Marquette University, Milwaukee, Wisconsin 53233
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- Winfield S. Sale
- Department of Cell Biology, Emory University School of Medicine, Atlanta, Georgia 30322
抄録
<jats:title>ABSTRACT</jats:title> <jats:p> Genetic and morphological studies have revealed that the radial spokes regulate ciliary and flagellar bending. Functional and biochemical analysis and the discovery of calmodulin in the radial spokes suggest that the regulatory mechanism involves control of axonemal protein phosphorylation and calcium binding to spoke proteins. To identify potential regulatory proteins in the radial spoke, in-gel kinase assays were performed on isolated axonemes and radial spoke fractions. The results indicated that radial spoke protein 2 (RSP2) can bind ATP and transfer phosphate in vitro. <jats:italic>RSP2</jats:italic> was cloned and mapped to the <jats:italic>PF24</jats:italic> locus, a gene required for motility. Sequencing revealed that <jats:italic>pf24</jats:italic> contains a point mutation converting the first ATG to ATA, resulting in only trace amounts of RSP2 and confirming the <jats:italic>RSP2</jats:italic> mapping. Surprisingly, the sequence does not include signature domains for conventional kinases, indicating that RSP2 may not perform as a protein kinase in vivo. However, the predicted RSP2 protein sequence contains Ca <jats:sup>2+</jats:sup> -dependent calmodulin binding motifs and a GAF domain, a domain found in diverse signaling proteins for binding small ligands including cyclic nucleotides. As predicted from the sequence, recombinant RSP2 binds calmodulin in a calcium-dependent manner. We postulate that RSP2 is a regulatory subunit of the radial spoke involved in localization of calmodulin for control of motility. </jats:p>
収録刊行物
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- Eukaryotic Cell
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Eukaryotic Cell 3 (1), 72-81, 2004-02
American Society for Microbiology
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詳細情報 詳細情報について
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- CRID
- 1361137046104775552
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- NII論文ID
- 30002293608
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- ISSN
- 15359786
- 15359778
- http://id.crossref.org/issn/15359778
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