Study of the effects of PRMT7-catalyzed methylation on the function and expression of the RNA-binding protein Alba20 in Leishmania major

• Grant number: 14/19400-1
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: March 01, 2015
• End date: October 09, 2015
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Angela Kaysel Cruz
• Grantee: Tiago Rodrigues Ferreira
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

Arginine methylation is a widely distributed post-translational modification. Protein arginine methyltransferase (PRMTs) enzymes, which catalize this modification, have been described in parasitic protozoa. Recently, our research group showed the expression profile of the PRMT7 enzyme from Leishmania major (LmjPRMT7) during promastigote development and its implication in parasite virulence. We observed that LmjPRMT7 associates to nine different RNA-binding proteins, including LmjAlba20, which is only arginine methylated in the presence of the LmjPRMT7 gene. The knockout of this gene revealed a dramatic alteration in LmjAlba20 expression pattern in stationary phase promastigotes. In this project, we propose to investigate a putative regulatory mechanism promoted by arginine methylation on LmjAlba20 affinity for specific transcripts and on the levels of this protein during L. major development. Initially, we will assess LmjPRMT7 methylation activity on LmjAlba20 in vitro, using heterologous protein. In the in vivo analyses, we will compare samples from wild-type L. major and null mutante ”lmjprmt7. The transcripts associated to myc-LmjAlba20 will be purified and identified by RNA sequencing (RNA-seq). The expression profile of methylated LmjAlba20 mimetic mutants will be evaluated in the different parasite stages. The proposed study will allow the investigation of a regulatory mechanism yet unexplored in tripanosomatids.

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)

CARNEIRO ALVES-FERREIRA, ELIZA VANESSA; FERREIRA, TIAGO RODRIGUES; WALRAD, PEGINE; KAYE, PAUL M.; CRUZ, ANGELA KAYSEL. Leishmania braziliensis prostaglandin F-2 alpha synthase impacts host infection. PARASITES & VECTORS, v. 13, n. 1, JAN 8 2020. (13/50219-9, 11/02040-4, 14/19400-1, 10/20597-3)

FERREIRA, TIAGO R.; DOWLE, ADAM A.; PARRY, EWAN; ALVES-FERREIRA, ELIZA V. C.; HOGG, KAREN; KOLOKOUSI, FOTEINI; LARSON, TONY R.; PLEVIN, MICHAEL J.; CRUZ, ANGELA K.; WALRAD, PEGINE B.. PRMT7 regulates RNA-binding capacity and protein stability in Leishmania parasites. Nucleic Acids Research, v. 48, n. 10, p. 5511-5526, JUN 4 2020. (15/13618-8, 14/19400-1, 14/50954-3)