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Infection and Immunity, October 2008, p. 4757-4763, Vol. 76, No. 10
0019-9567/08/$08.00+0     doi:10.1128/IAI.00527-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Role of Protein Kinase A in Trypanosoma cruzi{triangledown} ,{dagger}

Yi Bao,1 Louis M. Weiss,1,2 Vicki L. Braunstein,1 and Huan Huang1*

Department of Pathology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York 10461,1 Department of Medicine, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York 104612

Received 29 April 2008/ Returned for modification 27 May 2008/ Accepted 3 August 2008

Protein kinase A (PKA) is an important mediator of many signal transduction pathways that occur in eukaryotic cells, and it has been implicated as a regulator of stage differentiation in Trypanosoma cruzi. To evaluate the importance of the PKA catalytic subunit of T. cruzi (TcPKAc), a gene encoding a PKA inhibitor (PKI) containing a specific PKA pseudosubstrate, R-R-N-A, was subcloned into a pTREX vector and introduced into epimastigotes by electroporation. Expression of PKI has a lethal effect in this parasite. Similarly, a pharmacological inhibitor, H89, killed epimastigotes at a concentration of 10 µM. To understand the biology of PKA, identification of the particular substrates of this enzyme is essential. Using a yeast two-hybrid system, 38 candidates interacting with TcPKAc were identified. Eighteen of these were hypothetical proteins with unknown functions, while the others had putative or known functions. The entire open reading frames of eight genes presumably important in regulating T. cruzi growth, adaptation, and differentiation, including a type III PI3 kinase (Vps34), a putative PI3 kinase, a putative mitogen-activated extracellular signal-regulated kinase, a cyclic AMP (cAMP)-specific phosphodiesterase (PDEC2), a hexokinase, a putative ATPase, a DNA excision repair protein, and an aquaporin were confirmed to interact with TcPKAc in the yeast Saccharomyces cerevisiae under the highest stringency selection conditions, and PKA phosphorylated the recombinant proteins of these genes. Taken together, these findings demonstrate the importance of cAMP-PKA signaling in this organism.

* Corresponding author. Mailing address: Department of Pathology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461. Phone: (718) 430-2143. Fax: (718) 430-8543. E-mail: huangh{at}aecom.yu.edu

{triangledown} Published ahead of print on 11 August 2008.

{dagger} Supplemental material for this article may be found at http://iai.asm.org/.

Editor: J. F. Urban, Jr.

Infection and Immunity, October 2008, p. 4757-4763, Vol. 76, No. 10
0019-9567/08/$08.00+0     doi:10.1128/IAI.00527-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Bao, Y., Weiss, L. M., Hashimoto, M., Nara, T., Huang, H. (2009). Protein Kinase A Regulatory Subunit Interacts with P-Type ATPases in Trypanosoma cruzi. Am J Trop Med Hyg 80: 941-943 [Abstract] [Full Text]  


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