Evaluating the role of aconitase in the regulation of FeS synthesis genes in Caulobacter

• Grant number: 23/06833-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2023
• End date: June 30, 2024
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Marilis Do Valle Marques
• Grantee: João Pedro de Carvalho Pereira
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 21/10577-0 - Biology of Bacteria and Bacteriophages Research Center, AP.CEPID

Abstract

Aconitase is a protein best known for its enzymatic action of catalyzing the conversion reaction from citrate to isocitrate in the Krebs cycle when bound to a FeS group. Additionally, in other organisms such as Escherichia coli, this protein in the Apo form (without FeS) is also able to bind RNA, changing its half-life and translation. However, in C. crescentus (an alphaproteobacterium commonly used as a model organism for studying cell differentiation in prokaryotes), this protein is found in the RNA degradosome, an important structure for global regulation of gene expression in prokaryotes. Thus, this project aims to evaluate if the C. crescentus aconitase can bind RNAs and to which, and how it interferes with the regulation of these RNAs. For this purpose, the aconitase gene will be cloned with a FLAG tag in C. crescentus, the protein will be purified with the ligand RNAs, and these will be sequenced. The changes in the stability of these RNAs will be evaluated through RT-qPCR in cultures with normal iron concentrations and in the absence of iron.