The role of ac4C RNA modification in the Leishmania stages differentiation

• Grant number: 23/16672-0
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: April 01, 2024
• End date: March 31, 2028
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Nilmar Silvio Moretti
• Grantee: Ariely Barbosa Leite
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

Leishmania spp. is the etiological agent of leishmaniasis, a disease that affects humans and animals. During its life cycle, the parasite migrates from a vector to a vertebrate host facing distinct environmental conditions that require changes in various cellular processes, i.e., gene expression and metabolism, for its adaptation and survival. The mechanisms that govern Leishmania adaptation could involve, amongst others, chemical RNA modifications, i.e., N4-acetylcytidine (ac4C), to regulate gene expression and protein synthesis. Different chemical modifications have been identified in tRNAs, rRNAs, and mRNAs, known as epitranscriptome. The ac4C is catalyzed by the N-acetyltransferase 10 (NAT10) and depending on the position ac4C is added in mRNA could greatly impact the stability and/or the translation efficiency of each specific molecule. Considering that gene expression regulation happens mainly at the post-transcriptional level and the impact that ac4C can have on mRNA metabolism, we hypothesized that this modification might play an important role in Leishmania differentiation and adaptation throughout its life cycle. Preliminary data from our group identified a NAT10 orthologue and the presence of ac4C in different Leishmania species, i.e., Leishmania braziliensis and Leishmania tropica. To better understand the function of this modification in Leishmania, we will use different biochemistry and molecular biology techniques in this project, to: (i) Quantify the ac4C-modified RNA levels of the stages (procyclic, metacyclic and amastigote) of Leishmania; (ii) identify the ac4C-modified RNAs in the different forms of Leishmania by RIP-seq assays; (iii) characterize and identify NAT10 partners in Leishmania. This project is very innovative and will help to increase our knowledge of how Leishmania regulates gene expression regulation. Moreover, RNA modification is a very interesting and timely topic as emphasized by this year's Nobel Prize of Physiology and Medicine to Katalin Karikó and Drew Weissman for their remarkable accomplishments in this field, and any research in this topic will contribute to different areas.