Characterization of zinc-dependent lysine deacetylase 4 of Leishmania mexicana

• Grant number: 20/01434-8
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2020
• End date: December 31, 2021
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Nilmar Silvio Moretti
• Grantee: Myrna Victória Zanchetta Costa
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

Leishmaniasis are caused by diferente species of Leishmania spp. The disease has a clinical spectrum ranging from cutaneus and mucocutaneous lesions to visceral lesions. During its life cycle, Leishmania sp. transit between invertebrate and vertebrate hosts, facing different environmental changes, which require rapid adaptations of the parasite for its survival. Post- translational modifications, such as acetylation and phosphorylation, have been implicate in the regulation of several cellular processes. Our group recently described the acetylome of promastigote, metacyclic and amastigote forms of Leishmania mexicana, and found 336 differentially lysine acetylated sites (K-ac) in 253 proteins of procyclic; 550 K-ac in 304 proteins in the metacyclic and 349 K-ac in 225 proteins in the amastigote form. Knowing that protein acetylation levels seems to play an important role in the regulatory mechanisms of Leishmania sp. among its evolutionary forms, this project aims to characterize the zinc-dependent lysine deacetylase 4 (DAC4) of L. mexicana. Using CRISPR/Cas9 system, we generate single-knockout, null and fluorescent "tagged" cell lines of DAC4 protein. Thus, using the strains, we intend in this project: I) to evaluate the role of DAC4 in the proliferation processes of the procyclic forms of L. mexicana; II) evaluate the involvement of DAC4 in the differentiation from procyclic forms to metacyclic and amastigote forms; III) investigate the expression of DAC4 in the evolutionary forms of the parasite; IV) analyze the subcellular location of DAC4 in the metacyclic and amastigote parasite forms. With this project we intend to contribute to a better understanding of the role of protein acetylation in the regulation of essential processes for the proliferation and progression of L. mexicana infection. (AU)