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Bacterial transcription factors involved in metal homeostasis, oxidative stress and virulence: investigating how Chromobacterium violaceum switches from the environment to the host

Grant number: 21/06894-0
Support Opportunities:Research Grants - Young Investigators Grants - Phase 2
Duration: July 01, 2022 - June 30, 2027
Field of knowledge:Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
Principal Investigator:José Freire da Silva Neto
Grantee:José Freire da Silva Neto
Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated researchers: Eric Skaar ; Luis Eduardo Soares Netto ; Tie Koide
Associated research grant:12/20435-9 - Transcription factors of Chromobacterium violaceum: integrating signaling pathways, regulons and pathogenicity, AP.JP
Associated scholarship(s):23/09116-3 - Global response of Chromobacterium violaceum to metal limitation mediated by the host-defense protein calprotectin, BP.PD
23/03796-2 - Mechanisms of regulation and transport of manganese and their relationship with the virulence of Chromobacterium violaceum, BP.PD
23/02296-6 - Techniques of molecular biology and microbiology as support to study genes involved in metal homeostasis and virulence in Chromobacterium violaceum, BP.TT
+ associated scholarships 23/02350-0 - Maintenance of Galleria mellonella larvae for virulence assays with Chromobacterium violaceum, BP.TT
22/12782-2 - Determining the Zur regulon in Chromobacterium violaceum, BP.MS
22/05485-1 - Characterization of regulatory pathways and adhesins involved in biofilm formation and pathogenicity of Chromobacterium violaceum, BP.DR
20/15268-2 - Mechanisms of heme homeostasis in Chromobacterium violaceum, BP.DR
22/05740-1 - Role of a P-type ATPase in metal efflux in Chromobacterium violaceum, BP.IC
22/07859-6 - Techniques of molecular biology and microbiology as support to study genes involved in metal homeostasis and virulence in Chromobacterium violaceum, BP.TT - associated scholarships

Abstract

Bacteria have sophisticated mechanisms to survive in hostile environments. Bacterial pathogens must deal with harsh stress conditions inside the host, such as oxidative stress and limitation of the essential metals iron, zinc, and manganese. This project aims to unravel how the environmental opportunistic pathogen Chromobacterium violaceum coordinates adaptive responses of metal homeostasis and antioxidant defense with the expression of virulence factors to survive in the host. We will construct a transposon mutant library in C. violaceum and develop an alternative infection model using Galleria mellonella. The library will be screened in G. mellonella to discover novel C. violaceum virulence factors and on CAS plates to identify novel transcription factors involved in siderophore regulatory networks. RNA sequencing (RNA-seq) will be used to identify the C. violaceum responses to intracellular infection, metal limitation mediated by the host-defense protein calprotectin, and metal intoxication by heme and manganese. The regulons of several C. violaceum transcription factors involved with metal homeostasis, oxidative stress, and virulence will be determined by RNA-seq and ChIP-seq. Null mutant strains will be used to characterize (I) the role of transporters for the uptake or efflux of iron, heme, zinc, and manganese; (II) the role of glutathione and the GarAB system in metal and oxidative stress defense; (III) the role of the type VI secretion system (T6SS) in metal uptake; (IV) the role of transcription factors in biofilm formation and antibiotic resistance. Data obtained in this project will reveal bacterial strategies to overcome host nutritional immunity. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
ALVES, JULIA A.; LEAL, FERNANDA C.; PREVIATO-MELLO, MARISTELA; DA SILVA NETO, JOSE F.. A Quorum Sensing-Regulated Type VI Secretion System Containing Multiple Nonredundant VgrG Proteins Is Required for Interbacterial Competition in Chromobacterium violaceum. MICROBIOLOGY SPECTRUM, v. N/A, p. 15-pg., . (18/03979-1, 18/12461-6, 20/00259-8, 21/06894-0)

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