Characterization of a potential diguanylate cyclase from Pseudomonas aeruginosa

• Grant number: 22/08037-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2022
• End date: February 29, 2024
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Regina Lúcia Baldini
• Grantee: Nane Milene Sposito Almeida Pereira
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

The second messenger cyclic bis-(3',5')-di-guanosine monophosphate (c-di-GMP) influences the regulation of several bacterial processes involved in virulence, such as biofilm formation. The synthesis and degradation of c-di-GMP occur due to two classes of proteins: diguanylate cyclases (DGCs), with active GGDEF domains; and phosphodiesterases (PDEs) with EAL or HD-GYP domains. The opportunistic bacterium Pseudomonas aeruginosa harbors about 40 proteins with such domains, however, many of these proteins remain uncharacterized. Among these, is the PA14_04420 protein, which has the GGDEF domain and the PAS sensory domain, with its auxiliary domain PAC. Previous results have shown a putative interaction of PA14_04420 with proteins involved in cell division, cell wall formation, and lipopolysaccharide synthesis. Therefore, this project aims to evaluate the potential catalytic activity of diguanylate cyclase of the PA14_04420 protein by expressing and purifying the recombinant protein and also evaluating concentrations of c-di-GMP produced in vivo, and verifying the enzymatic activity in vitro. Thus, the characterization of PA14_04420 may help to better understand the roles of c-di-GMP signaling pathways, which may provide a new target for therapeutic development against bacterial infections.(AU)