The protozoan parasite Trypanosoma cruzi, the causative agent of Chagas' disease, has emerged as a HIV/AIDS-related opportunistic infection. Current anti-T. cruzi agents are highly toxic, and there is no effective treatment for chronic Chagas' disease. Improvement in our understanding of the molecular signaling responsible for its differentiation may lead to effective inhibitors of these processes providing new and novel therapeutic approaches. Differentiation of T. cruzi involves cAMP, but the pathway in which it is acting remains unknown. It is likely, that as in other eukaryotes, cAMP-dependent protein kinase (PKA) is a major signal transduction pathway controlling cell differentiation. The PKA catalytic subunit (TcPKA-C) gene and the PKA regulatory subunit gene (TcPKA-R) of T. cruzi were cloned by our laboratory and data indicates that the PKA catalytic and regulatory subunits are developmentally regulated. These data support the hypothesis that PKA activity is important in T. cruzi stage differentiation. We now plan to study the functions of TcPKA in differentiation. The effects of both overexpression of TcPKA with a constitutively active TcPKAC and blockade of TcPKA with a PKI clone, a dominant negative inhibitory TcPKA-R as well as conditional knockout of TcPKA-C on differentiation and proliferation will be studied in transfected parasites. Mechanisms of intracellular targeting of TcPKA-R in T. cruzi will be examined by deletion analysis of Nterminus of TcPKA-R and by analyzing the post-translational modifications of TcPKA-R with mass spectrometry. To further delineate the components of this pathway, we will identify the TcPKA downstream interacting molecules, using both yeast two-hybrid assays and proteomic approaches. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI058893-01A2
Application #
6946961
Study Section
Special Emphasis Panel (ZRG1-AARR-A (04))
Program Officer
Rogers, Martin J
Project Start
2005-04-01
Project End
2010-03-31
Budget Start
2005-04-01
Budget End
2006-03-31
Support Year
1
Fiscal Year
2005
Total Cost
$365,831
Indirect Cost
Name
Albert Einstein College of Medicine
Department
Pathology
Type
Schools of Medicine
DUNS #
110521739
City
Bronx
State
NY
Country
United States
Zip Code
10461
Mukherjee, Shankar; Sadekar, Nikaeta; Ashton, Anthony W et al. (2013) Identification of a functional prostanoid-like receptor in the protozoan parasite, Trypanosoma cruzi. Parasitol Res 112:1417-25
Ma, Yan Fen; Weiss, Louis M; Huang, Huan (2012) A method for rapid regulation of protein expression in Trypanosoma cruzi. Int J Parasitol 42:33-7
Huang, Huan (2011) Signal transduction in Trypanosoma cruzi. Adv Parasitol 75:325-44
Huang, Huan; Ma, Yan Fen; Bao, Yi et al. (2011) Molecular cloning and characterization of mitogen-activated protein kinase 2 in Toxoplasma gondii. Cell Cycle 10:3519-26
Taylor, Martin C; Huang, Huan; Kelly, John M (2011) Genetic techniques in Trypanosoma cruzi. Adv Parasitol 75:231-50
Bao, Yi; Weiss, Louis M; Ma, Yan Fen et al. (2010) Molecular cloning and characterization of mitogen-activated protein kinase 2 in Trypanosoma cruzi. Cell Cycle 9:2888-96
Nagajyothi, Fnu; Zhao, Dazhi; Machado, Fabiana S et al. (2010) Crucial role of the central leptin receptor in murine Trypanosoma cruzi (Brazil strain) infection. J Infect Dis 202:1104-13
Bao, Yi; Weiss, Louis M; Ma, Yan Fen et al. (2010) Protein kinase A catalytic subunit interacts and phosphorylates members of trans-sialidase super-family in Trypanosoma cruzi. Microbes Infect 12:716-26
Bao, Yi; Weiss, Louis M; Hashimoto, Muneaki et al. (2009) Protein kinase A regulatory subunit interacts with P-Type ATPases in Trypanosoma cruzi. Am J Trop Med Hyg 80:941-3
Villalta, Fernando; Scharfstein, Julio; Ashton, Anthony W et al. (2009) Perspectives on the Trypanosoma cruzi-host cell receptor interactions. Parasitol Res 104:1251-60

Showing the most recent 10 out of 13 publications