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