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First published online 13 September 2005
doi: 10.1242/jcs.02550

Journal of Cell Science 118, 4333-4341 (2005)
Published by The Company of Biologists 2005
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Research Article

Direct involvement of the isotype-specific C-terminus of ß tubulin in ciliary beating

Julia Vent1,*, Todd A. Wyatt2, D. David Smith1, Asok Banerjee3, Richard F. Ludueña3, Joseph H. Sisson2 and Richard Hallworth1,{ddagger}

1 Department of Biomedical Sciences, Creighton University, Omaha, Nebraska 68178, USA
2 Department of Internal Medicine, Section of Pulmonary, Critical Care and Sleep Medicine, University of Nebraska Medical Center, Omaha, Nebraska, USA
3 Department of Biochemistry, University of Texas Health Science Center at San Antonio, San Antonio, Texas, USA

{ddagger} Author for correspondence (e-mail: hallw{at}creighton.edu)

Accepted 17 June 2005

In previous studies in Drosophila, Nielsen et al. hypothesized that the ß tubulin C-terminal axonemal motif `EGEFXXX', where X is an acidic amino acid, is required for ciliary function and assembly (Nielsen et al., 2001, Curr. Biol. 11, 529-533). This motif is present in some but not all mammalian ß tubulin isotypes. We therefore investigated whether this motif is important in ciliary function in mammals. In a preparation of isolated, ATP-reactivated bovine tracheal cilia, we found that monoclonal antibodies directed against the C-terminus of ßI, ßIV and ßV tubulin blocked ciliary beating in a concentration dependent manner. Antibodies against other epitopes of ß tubulin were ineffective, as were antibodies against {alpha} tubulin. Peptides consisting of the axonemal motif and motif-like sequences of these isotypes blocked ciliary beating. These results suggest that the axonemal motif sequences of ßI, ßIV and ßV tubulin are essential for ciliary function. Peptides consisting of corresponding C-terminal sequences in {alpha} tubulin isotypes were also ineffective in blocking ciliary beating, which suggests that the C-terminus of {alpha} tubulin is not directly involved in cilia function in mammals.

Key words: Axonemal function, ß Tubulin isotype, Cilia, Ciliary beat frequency


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© The Company of Biologists Ltd 2005