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J. Biol. Chem., Vol. 280, Issue 16, 16235-16243, April 22, 2005

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A Lipid-modified Phosphoinositide-specific Phospholipase C (TcPI-PLC) Is Involved in Differentiation of Trypomastigotes to Amastigotes of Trypanosoma cruzi^*

Michael Okura, Jianmin Fang, Maria Laura Salto, Randall S. Singer¶, Roberto Docampo, and Silvia N. J. Moreno||

From the Department of Pathobiology and Center for Zoonoses Research, University of Illinois at Urbana-Champaign, Urbana, Illinois 61802 and the Center for Tropical and Global Emerging Diseases and Department of Cellular Biology, University of Georgia, Athens, Georgia 30602

The phosphoinositide-specific phospholipase C (PI-PLC) is an important component of the inositol phosphate/diacylglycerol signaling pathway. A newly discovered Trypanosoma cruzi PI-PLC (TcPI-PLC) is lipid modified in its N terminus, targeted to its plasma membrane, and believed to play a role in differentiation of the parasite because its expression increases during the differentiation of trypomastigote to amastigote stages. To determine whether TcPI-PLC is involved in this differentiation step, antisense inhibition using phosphorothioate-modified oligonucleotides, and overexpression of the gene were performed. Antisense oligonucleotide-treated parasites showed a reduced rate of differentiation in comparison to controls, as well as accumulation of intermediate forms. Overexpression of TcPI-PLC led to a faster differentiation rate. In contrast, overexpression of a mutant TcPI-PLC that lacked the lipid modification at its N terminus did not affect the differentiation rate. Therefore, TcPI-PLC is involved, when expressed in the plasma membrane, in the differentiation of trypomastigotes to amastigotes, an essential step for the intracellular replication of these parasites.

Received for publication, December 23, 2004

^* This work was supported in part by the American Heart Association, Midwest Affiliate, National Institutes of Health Grant AI-48039 (to S. N. J. M.), and by a facility constructed in part with support from Research Facility Improvement Grant Number C06 RR16515-01 from the National Center for Research Resources, National Institutes of Health. The costs of publication of this article were defrayed in part by the payment of page charges. This 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 Veterinary and Biomedical Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul, MN 55108.

^|| To whom correspondence should be addressed: Center for Tropical and Global Emerging Diseases and Dept. of Cellular Biology, University of Georgia, Athens, GA 30602. Tel.: 706-542-4736; Fax: 706-542-3582; E-mail: s-moreno{at}cb.uga.edu.

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