Identification and characterization of actin-binding proteins (ABPs) of the Apicomplexan parasite Neospora caninum

• Grant number: 12/22772-2
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: February 01, 2013
• End date: January 31, 2017
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Ana Patricia Yatsuda Natsui
• Grantee: Luciana Baroni
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated scholarship(s): 15/04258-8 - Actin-binding proteins (ABPs) from Neospora caninum: role of actin-depolymerizing factor (NcADF) and cyclase-associated protein (NcCAP) in actin dynamics, BE.EP.DR

Abstract

Neospora caninum is an Apicomplexan protozoan that infects, among a whole range of intermediate hosts, bovine where it is emerging as a relevant cause of reproductive problems and abortion in dairy and beef cattle. As obligatory intracellular organisms, Apicomplexan parasites use their own active locomotion system to move and invade host cells. This mechanism is driven by the actin/myosin motor known as gliding motility, localized between the plasma and the inner membrane complex. Actin has a central role in gliding motility of Apicomplexan organisms and presents a unique dynamic, having a turnover tightly regulated by actin binding proteins (ABPs). ABPs have ability to bind actin and act in its polymerization and depolymerization. This project aims to identifying and characterizing N. caninum ABPs. In order to achieve that, tools will be developed such as specific antibody against a recombinant actin epitope between aminoacids 201 and 310 of N. caninum actin (NcAct201-310) and an antibody against recombinant N. caninum ABP (NcABP). The ABP will be chosen after an in silico analysis of NcABPs, cloned and expressed in this project. Other tools to be made include the ABP knockout in N. caninum tachyzoites and tachyzoites transfected with YFP-labeled NcABP or under control of Tet expression repressor system (TetR-tetO). After the achievement of these tools, tests will be conducted to characterized and evaluate the NcABP function using wild, knockout and transfected tachyzoites in confocal and electronic microscopy. Then, these tachyzoites will be used to perform 1 D and 2 D SDS-PAGE, followed by western blot and identification of spots and bands by mass spectrometry. Finally, the polymerization of rabbit muscle actin with or without NcABP will also be evaluated by fluorometric technique.

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)

BARONIL, LUCIANA; POLLO-OLIVEIRA, LETICIA; HECK, ALBERT J. R.; MAARTEN ALTELAAR, A. F.; YATSUDA, ANA PATRICIA. Actin from the apicomplexan Neospora caninum (NcACT) has different isoforms in 2D electrophoresis. Parasitology, v. 146, n. 1, p. 33-41, JAN 2019. (12/22772-2)

BARONI, LUCIANA; PEREIRA, LUIZ M.; MACIVER, SUTHERLAND K.; YATSUDA, ANA P.. Functional characterisation of the actin-depolymerising factor from the apicomplexan Neospora caninum (NcADF). Molecular and Biochemical Parasitology, v. 224, p. 26-36, SEP 15 2018. (15/04258-8, 12/22772-2)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

BARONI, Luciana. Identification and characterization of actin binding proteíns (ABPs) from the apicomplexan Neospora caninum. 2017. Doctoral Thesis - Universidade de São Paulo (USP). Faculdade de Ciências Farmacêuticas de Ribeirão Preto (PCARP/BC) Ribeirão Preto.