Chromobacterium violaceum transcriptome in cell cultures and during infection in mouse

• Grant number: 18/12461-6
• Support type: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): February 01, 2019
• Effective date (End): March 31, 2022
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal researcher: José Freire da Silva Neto
• Grantee: Maristela Previato Mello
• Home Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

During infection occurs modulation of gene expression in both pathogen and host. Chromobacterium violaceum is an opportunistic pathogen that causes severe infections characterized by septicemia that can lead to death. However, no study characterized in a global way alterations in gene expression during pathogen-host interaction in mammalian infection caused by C. violaceum. Thus, we propose to identify the global transcriptional changes during C. violaceum infection in cell cultures and animal tissue in vivo using RNA sequencing (RNA-Seq). For this, we will characterize C. violaceum infection in cell cultures of hepatocytes and macrophages using transmission electron microscopy. Then, we will sequence the C. violaceum RNA during infection in the liver, hepatocyte and mouse macrophage, as well as sequence the macrophage RNA infected with this bacterium using the RNA-Seq and Dual RNA-Seq techniques. We will perform a global analysis of these transcripts focusing on the identification of genes for virulence factors and transcription regulators, mainly regulators of the MarR family, which are already targets of in vitro studies and with role in virulence seen by our group. The differentially expressed genes selected will be validated by qRT-PCR and analyzed for their role in virulence. Thus, the transcriptomes obtained and analyzed in this work under infective conditions should contribute to the discovery of the regulatory networks involved in the in vivo expression of virulence determinants of C. violaceum and host defense pathways.