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Vol. 16, Issue 7, 3247-3259, July 2005

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ACT-5 Is an Essential Caenorhabditis elegans Actin Required for Intestinal Microvilli Formation

A. J. MacQueen ^* , J. J. Baggett ^* , N. Perumov ^*, R. A. Bauer ^*, T. Januszewski , L. Schriefer ^||, and J. A. Waddle ^* 

^* Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390; Molecular and Cellular Imaging Facility, University of Texas Southwestern Medical Center, Dallas, TX 75390; and ^|| Department of Genetics, Washington University School of Medicine, St. Louis, MO 63110

Submitted December 10, 2004; Revised March 30, 2005; Accepted April 25, 2005
Monitoring Editor: Susan Strome

Investigation of Caenorhabditis elegans act-5 gene function revealed that intestinal microvillus formation requires a specific actin isoform. ACT-5 is the most diverged of the five C. elegans actins, sharing only 93% identity with the other four. Green fluorescent protein reporter and immunofluorescence analysis indicated that act-5 gene expression is limited to microvillus-containing cells within the intestine and excretory systems and that ACT-5 is apically localized within intestinal cells. Animals heterozygous for a dominant act-5 mutation looked clear and thin and grew slowly. Animals homozygous for either the dominant act-5 mutation, or a recessive loss of function mutant, exhibited normal morphology and intestinal cell polarity, but died during the first larval stage. Ultrastructural analysis revealed a complete loss of intestinal microvilli in homozygous act-5 mutants. Forced expression of ACT-1 under the control of the act-5 promoter did not rescue the lethality of the act-5 mutant. Together with immuno-electron microscopy experiments that indicated ACT-5 is enriched within microvilli themselves, these results suggest a microvillus-specific function for act-5, and further, they raise the possibility that specific actins may be specialized for building microvilli and related structures.

The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).

Present address: Molecular, Cellular, and Developmental Biology Department, Yale University, New Haven, CT 06511

Present address: Biology Department, Southern Methodist University, Dallas, TX 75275.

Address correspondence to: J. A. Waddle (jwaddle{at}mail.smu.edu).

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