Analysis of LmjPRMT7 function during Leishmania major development

• Grant number: 13/22222-5
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: January 01, 2014
• End date: June 30, 2014
• Field of knowledge: Biological Sciences - Biochemistry - Biochemistry of Microorganisms
• Principal Investigator: Angela Kaysel Cruz
• Grantee: Tiago Rodrigues Ferreira
• Supervisor: Deborah Frances Smith
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Institution abroad: University of York, England
• Associated to the scholarship: 10/01129-9 - Investigation of the function of the arginine methyltransferase LmjPRMT7 on the control of gene expression in Leishmania major, BP.DR

Abstract

Arginine methylation is an evolutionarily conserved protein post-translational modification. Protein arginine methyltransferases (PRMTs) have been described in several organisms from animals to early branching protozoa. We have recently identified several binding partners and putative LmjPRMT7 substrates in Leishmania major, including RNA binding proteins (RBP). LmjPRMT7-/- parasites exhibit increased infectivity both in vitro and in vivo while mice infected with LmjPRMT7 overexpressing parasites displayed reduced lesion size progression. To further investigate LmjPRMT7 function, we propose this joint collaboration with Prof. Deborah Smith and Dr. Pegine Walrad, York University, UK. We will generate co-transfectants of tagged LmjPRMT7 and putative substrates, using the previously generated knockout parasites. Their expertise on parasite differentiation and characterization of tripanossomatid RNA binding proteins will enhance the quality of our work. To validate previously identified LmjPRMT7 interactions, we will use immunoprecipitations and in vitro methylation assays. We will complement LmjPRMT7-/- parasites with a tagged version of this enzyme regulated either by its native 3'UTR or by the 3'UTR of SHERP (small hydrophilic ER-associated protein) to focus LmjPRMT7 expression to stationary phase and check for distinct phenotypes. We will analyze the RNA binding properties of RBP substrates in absence or presence of arginine methylation. This study will provide means to characterize LmjPRMT7 protein substrates and to further analyze its function in L. major. (AU)