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Originally published In Press as doi:10.1074/jbc.M200873200 on March 4, 2002

J. Biol. Chem., Vol. 277, Issue 20, 17580-17588, May 17, 2002
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Trypanosoma brucei FLA1 Is Required for Flagellum Attachment and Cytokinesis*

Douglas J. LaCountDagger , Brian Barrett, and John E. Donelson§

From the Department of Biochemistry, University of Iowa, Iowa City, Iowa 52242

The single flagellum of the protozoan parasite Trypanosoma brucei is attached along the length of the cell body by a complex structure that requires the FLA1 protein. We show here that inhibition of FLA1 expression by RNA interference in procyclic trypanosomes causes flagellar detachment and prevents cytokinesis. Despite being unable to divide, these cells undergo mitosis and develop a multinucleated phenotype. The Trypanosoma cruzi FLA1 homolog, GP72, is unable to complement either the flagellar detachment or cytokinesis defects in procyclic T. brucei that have been depleted of FLA1 by RNA interference. Instead, GP72 itself caused flagellar detachment when expressed in T. brucei. In contrast to T. brucei cells depleted of FLA1, procyclic T. brucei expressing GP72 continued to divide despite having detached flagella, demonstrating that flagellar attachment is not absolutely necessary for cytokinesis. We have also identified a FLA1-related gene (FLA2) whose sequence is similar but not identical to FLA1. Inhibition of FLA1 and FLA2 expression in bloodstream T. brucei caused flagellar detachment and blocked cytokinesis but did not inhibit mitosis. These experiments demonstrate that the FLA proteins are essential and suggest that in procyclic T. brucei, the FLA1 protein has separable functions in flagellar attachment and cytokinesis.

* This work was supported by Grants AI10512, AI40591, and DK25295 from the National Institutes of Health.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Present address: Dept. of Genome Sciences, University of Washington, Box 357730, Seattle, WA 98195-7730.

§ To whom correspondence should be addressed: Tel.: 319-335-7934; Fax: 319-353-4204; E-mail: john-donelson@uiowa.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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