Heterologous Expression of a PilZ Homolog for Studying c-di-GMP Signaling in Leptospira interrogans

• Grant number: 23/07366-2
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: September 01, 2023
• End date: December 31, 2024
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Cristiane Rodrigues Guzzo Carvalho
• Grantee: Cauê Augusto Boneto Gonçalves
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Bacteria of the genus Leptospira are renowned for causing a disease called Leptospirosis, which poses a substantial public health problem and is grossly neglected. Consequently, due to both the challenges associated with manipulating these bacteria in a laboratory environment and the lack of attention given to them, many fundamental aspects remain inadequately explored. One such aspect pertains to intracellular signaling pathways, which constitute the focus of this study through the heterologous expression of PilZ via the LIC_10128 gene.Furthermore, this protein harbors a binding site for c-di-GMP, which underlies diverse phenotypic traits observed in gram-negative bacteria. Therefore, the structure of the target protein is determined via crystallization after cloning and expression in E. coli. Additionally, an interaction test with c-di-GMP is performed, enabling the investigation of LIC_10128's role in c-di-GMP binding, and the endeavor to ascertain its three-dimensional structure in its apo form as well as in complex with c-di-GMP. Notably, as the PilZ domain protein lacks a documented functional description in the literature, any insights gained from this project will contribute to enhancing our comprehension of the c-di-GMP signaling network.