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

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

Julia Vent^1,*, Todd A. Wyatt², D. David Smith¹, Asok Banerjee³, Richard F. Ludueña³, Joseph H. Sisson² and Richard Hallworth^1,

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

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 ß_I, ß_IV and ß_V tubulin blocked ciliary beating in a concentration dependent manner. Antibodies against other epitopes of ß tubulin were ineffective, as were antibodies against 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 ß_I, ß_IV and ß_V tubulin are essential for ciliary function. Peptides consisting of corresponding C-terminal sequences in tubulin isotypes were also ineffective in blocking ciliary beating, which suggests that the C-terminus of 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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