Signaling pathways involved in the interaction of Trypanosoma cruzi with Extracellular Matrix isolated from distinct cell lines

• Grant number: 17/19854-0
• Support Opportunities: Regular Research Grants
• Start date: December 01, 2017
• End date: August 31, 2020
• Field of knowledge: Biological Sciences - Biochemistry - Biochemistry of Microorganisms
• Principal Investigator: Maria Julia Manso Alves
• Grantee: Maria Julia Manso Alves
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Walter Colli

Abstract

S-nitrosylation and phosphorylation are reversible post-translational modifications that have been implicated in the regulation of protein function. We described recently that S-nitrosylation and phosphorylation of proteins occur in T. cruzi trypomastigotes during their interaction with ECM, an obligatory step in the invasion process. Unpublished data (in collaboration with G. Palmisano) showed that proteins from the cytoskeleton (paraflagellar rod protein, ± and ²-tubulin), phosphatases, kinases and ribosomal proteins, among others, are S-nitrosylated and/or phosphorylated. This project has two main goals: to determine the influence of the ECM composition on the post-translational modification of T. cruzi proteins and to search for signaling pathways as a consequence of the adhesion to ECM. For these purposes, ECMs will be prepared from three cell lines already selected by their high or low capacity of adhesion to or invasion by T. cruzi. The composition of isolated ECMs will be determined by proteomic analysis. To get insights on the signaling pathway in ECMs-treated trypomastigotes, NO synthase activity, NO production, cGMP, cAMP, kinases activities (AMPK, PKA, MAPK, GSK-3 and PIK3), SNO and phosphorylation profiles in the presence of inhibitors of NO synthase, phosphodiesterase, protein kinase and phosphatases will be analyzed in a time period. To determine the possible role of AMPK on the carbohydrate metabolism, the glucose transport will be measured. In addition, total protein synthesis and the levels of S-nitrosylation, sumoylation and ubiquitinylation of the paraflagellar rod protein will be determined in the ECMs-treated trypomastigotes under the same experimental conditions described above.This project intends to understand better the signaling pathways activated in trypomastigotes during their adhesion to ECMs isolated from different cell lines, which may contribute to the understanding of T. cruzi invasion mechanisms. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

COUTINHO, JOAO V. P.; ROSA-FERNANDES, LIVIA; MULE, SIMON NGAO; DE OLIVEIRA, GILBERTO SANTOS; MANCHOLA, NUBIA CAROLINA; SANTIAGO, VERONICA FEIJOLI; COLLI, WALTER; WRENGER, CARSTEN; MANSO ALVES, MARIA JULIA; PALMISANO, GIUSEPPE. The thermal proteome stability profile of Trypanosoma cruzi in epimastigote and trypomastigote life stages. JOURNAL OF PROTEOMICS, v. 248, SEP 30 2021. (18/13283-4, 18/15549-1, 14/06863-3, 17/03966-4, 15/26722-8, 17/19854-0, 17/04032-5, 18/18257-1)