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MBC in Press, published online ahead of print May 4, 2005
Mol. Biol. Cell 10.1091/mbc.E04-12-1061

A more recent version of this article appeared on July 1, 2005
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Submitted on December 10, 2004
Revised on March 30, 2005
Accepted on April 25, 2005

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

A. J. MacQueen,*{dagger} J. J. Baggett,*{ddagger} N. Perumov,* R. A. Bauer,* T. Januszewski,{sect} L. Schriefer,|| and J. A. Waddle*{ddagger}

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

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. GFP reporter and immunofluoresence 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 appeared 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. Taken together with immuno-EM experiments that indicated ACT-5 is enriched within microvilli themselves, these results suggest a microvillus - specific function for act-5 and, further, raise the possibility that specific actins may be specialized for building microvilli and related structures.

Present addresses: {dagger}Molecular, Cellular and Developmental Biology Department, Yale University, New Haven, CT 06511; {ddagger}Biology Department, Southern Methodist University, Dallas, TX 75275. J. A. Waddle (jwaddle{at}mail.smu.edu)




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