CRK3 (Cdc2 Related Kinase): Uncovering modulation pathways linked to cell cycle and metacyclogenesis in Trypanosoma cruzi

• Grant number: 22/05264-5
• Support Opportunities: Scholarships in Brazil - Master
• Start date: November 01, 2022
• End date: July 31, 2024
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Simone Guedes Calderano
• Grantee: Bruno Alves Santarossa
• Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
• Associated research grant: 20/00694-6 - How DNA replication contributes for the success of infection caused by Trypanosoma cruzi, AP.TEM

Abstract

Trypanosoma cruzi is a clinically relevant protozoan, causative agent of Chagas disease. Its endemic characteristic to emergent countries is an important contextualization to understand why it still is, proportionally to other model eukaryotes, very little understood. During its life cycle, T. cruzi transit between an invertebrate vector and a mammalian host and in these environments it presents different life forms: epimastigote (vector) and amastigotes (host) which are able to replicate then they differentiate in trypomastigote form which is able to infect but cannot replicate. These various changes that the parasite goes through indicate that there is a high degree of complexity of regulation of cellular mechanisms, in which control of progression of cell cycle is specially important in replication halt and in its replicative stages. Furthermore, replication is also a factor involved in resistance to treatment with benznidazole, since it was observed (in vitro and in vivo) that resistant parasites present low replication rate. In model eukaryotes pairs kinases CDKs (cyclin dependent kinases)-cyclins are responsible for cell cycle progression and regulation, they also take part in cellular replication halt in situations where there is lack of nutrients and mitogenic stimuli. In T. cruzi homologous genes to CDK1, named CRKs (Cdc2 related kinases) and in between these genes the enzyme translated by CRK3 is involved in cell cycle regulation and in the transition G2/M. Thus, it is proposed to explore what role the protein expressed by the CRK3 gene has in cell cycle regulation and metacyclogenesis. Utilizing techniques such as genetic editing by CRISPR/Cas9 and TurboID, it will be possible to identify the repertory of molecules that interact with CRK3 in different conditions (replicative epimastigotes and in metacyclogenesis), in such manner that establishment of CRK3 role in cell cycle regulation and quiescence entrance will be possible.