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Contractile eukaryotic flagella: Centrin is involved

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Summary

Isolated transverse flagella ofPeridinium inconspicuum (Dinophyceae) undergo a rapid Ca2+-induced (50μM Ca2+) contraction in the absence of exogenous ATP. Longitudinal flagella from the same species do not contract under these conditions. Contraction leads to a supercoiling of the axoneme and a shortening of the paraxonemal fiber that accompanies the axoneme over most of its length. Using a polyclonal antibody generated against centrin, a 20 kDa Ca2+-modulated contractile protein of striated flagellar roots of the green flagellateTetraselmis striata, we have found that the paraxonemal fiber in transverse flagella of three taxa ofDinophyceae is immunoreactive by indirect immunofluorescence. The localization of the antigen in the paraxonemal fiber of transverse flagella was confirmed by two-colour double immunofluorescence using monoclonal mouse-anti-β-tubulin for identification of the axoneme. No structure was immunoreactive to anticentrin in the longitudinal flagella of all taxa. Electrophoretic and immunoblot analysis of isolated flagella ofP. inconspicuum show that the antigen is a 21 kDa protein, indicating that it is either centrin or a closely related protein. We conclude that centrin confers contractility to the transverse flagellum of dinoflagellates and possibly to other contractile eukaryotic flagella.

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Abbreviations

ASP-H:

artificial seawater medium with Hepes-buffer

BSA:

bovine serum albumine

DTT:

dithiothreitol

EGTA:

ethylene glycol bis(2-amino-ethylether)tetraacetic acid

FITC:

fluorescein isothiocyanate

MT:

buffer microtubule stabilizing buffer

PBS:

phosphate buffered saline

SDS:

sodium dodecyl sulfate

TLCK:

Nα-p-tosyl-l-lysine chloromethyl ketone

TRITC:

tetramethylrhodamine isothiocyanate

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  1. J. Otten

    Present address: , Dorpsstraat 135, NL-6871, AG Renkum, Netherlands

Authors and Affiliations

  1. Botanisches Institut, Westfälische Wilhelms-Universität, Münster

    I. Höhfeld, J. Otten & M. Melkonian

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  1. I. Höhfeld
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  2. J. Otten
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  3. M. Melkonian
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Höhfeld, I., Otten, J. & Melkonian, M. Contractile eukaryotic flagella: Centrin is involved. Protoplasma 147, 16–24 (1988). https://doi.org/10.1007/BF01403874

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