Investigation of the structure-function relationship of Aedes aegypti proteins RPAP3, Pih1D1, WDR92 and their role in insect development

• Grant number: 24/01429-5
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: June 01, 2024
• End date: February 29, 2028
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Carlos Henrique Inacio Ramos
• Grantee: Larissa Machado Antonio
• Host Institution: Instituto de Química (IQ). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated research grant: 17/26131-5 - The chaperome: study of the relationship of the structure of its components and the maintenance of proteostasis, AP.TEM

Abstract

The R2TP complex (Rvb1-Rvb2-Tah1/RPAP3-Pih1/Pih1D1) is involved in the assembly and stabilization of macromolecular complexes associated with several pathways essential for cell growth and proliferation. More specifically, one of the components proteins of this complex, RPAP3, was shown to be essential for the development and survival of Drosophila melanogaster fruit fly larvae. Interestingly, this protein is not essential for yeast and humans, which suggests biological safety for its use as a molecular target to control mosquitoes, vectors of such important diseases in the national context. In the present project, we intend to structurally and functionally characterize the RPAP3, Pih1D1 and WDR92 proteins from Aedes aegypti, to shed light on their conformations and functions. Aiming to evaluate the importance of these proteins for the development and survival of the mosquito, we will perform analysis of spatial (in situ) and relative gene expression (qRT-PCR) at all stages of development, as well as gene silencing experiments using RNA interference and deletion by CRISPR/Cas9 of the rpap3, pih1d1 and wdr92 genes. We also aim to identify the formation of the R2TP complex in vivo through the co-localization of proteins in mosquito histological sections. Finally, we hope to identify compounds that interact with RPAP3, Pih1D1 and WDR92, destabilizing these proteins and, therefore, could be potential candidates for compounds to inhibit mosquito development at the larval stage. With this project we expect to publish at least two publications in indexed journals and contribute to the development of a product with commercial potential for controlling this vector.