Deconstructing complexity in the regulatory network for biofilm formation in gram-negative bacteria

• Grant number: 18/04810-0
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: August 01, 2018
• End date: May 31, 2023
• Field of knowledge: Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
• Principal Investigator: María Eugenia Guazzaroni
• Grantee: Ananda Sanches Medeiros
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 12/22921-8 - Synthetic biology approaches for deciphering the logic of signal integration in complex bacterial promoters, AP.JP
• Associated scholarship(s): 21/07728-6 - A high-throughput approach to understand biofilm promoter architecture under different growth conditions, BE.EP.DD

Abstract

Biofilms are complexes structures composite by different or similar bacteria that form communities, enclosed into a polymeric extracellular matrix (EPS), constituted by polysaccharides, proteins and nucleic acids. These structures are very present on the natural environment and exert an important role on ecologic and human life. Their formation regulatory networks are composite by environment and intracellular stimuli integration, able to drastically alter the bacterial gene expression, converting their motility behavior to a sessile. In this way, to understand better this genetic conversion is extremely important. Therefore, this project aims to investigate the role of the Global Transcription Factors Fis, IHF, CRP, FNR, ArcA, NarL and Lrp on the promoters of important biofilm regulators (such as rpoS, rpoE, csgD, cpxR, flhD, fliA, yeaJ, ompR, adrA and rpoD). This will be done under different experimental conditions, as pH variation, nutrients availability, temperature changes. Besides that, we will investigate the role of the regulatory regions 5'UTR aiming to understand on an integrated way the effect of post-transcriptional mechanism on the biofilm regulatory network. For this, we will analyze the gene expression variations levels, observing the GFP production and gene expression differential profile, using RNAseq. Besides evaluate the global TFs influences on the general biofilm gene expression, we will evaluate their functions on the three biofilm composition zones, utilizing confocal fluorescence microscopy and microdissection followed by RNA sequencing for verify the gene expression differences between the three zones. (AU)

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)

AMARELLE, VANESA; SANCHES-MEDEIROS, ANANDA; SILVA-ROCHA, RAFAEL; GUAZZARONI, MARIA-EUGENIA. Expanding the Toolbox of Broad Host-Range Transcriptional Terminators for Proteobacteria through Metagenomics. ACS SYNTHETIC BIOLOGY, v. 8, n. 4, p. 647-654, APR 2019. (18/04810-0, 15/04309-1)

OLIVEIRA MONTEIRO, LUMMY MARIA; SANCHES-MEDEIROS, ANANDA; WESTMANN, CAUA ANTUNES; SILVA-ROCHA, RAFAEL. Unraveling the Complex Interplay of Fis and IHF Through Synthetic Promoter Engineering. FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY, v. 8, JUN 18 2020. (18/04810-0, 12/22921-8, 16/05472-6, 16/19179-9, 17/50116-6)

OLIVEIRA MONTEIRO, LUMMY MARIA; ARRUDA, LETICIA MAGALHAES; SANCHES-MEDEIROS, ANANDA; MARTINS-SANTANA, LEONARDO; DE FATIMA ALVES, LUANA; DEFELIPE, LUCAS; GUSTAVO TURJANSKI, ADRIAN; GUAZZARONI, MARIA-EUGENIA; DE LORENZO, VICTOR; SILVA-ROCHA, RAFAEL. Reverse Engineering of an Aspirin-Responsive Transcriptional Regulator in Escherichia coli. ACS SYNTHETIC BIOLOGY, v. 8, n. 8, p. 1890-1900, AUG 2019. (16/19179-9, 17/17924-1, 16/06323-4, 18/04810-0, 15/04309-1, 12/22921-8)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

MEDEIROS, Ananda Sanches. Accessing transcriptional regulation of Escherichia coli biofilm formation using promoter and genomic libraries. 2023. Doctoral Thesis - Universidade de São Paulo (USP). Faculdade de Medicina de Ribeirão Preto (PCARP/BC) Ribeirão Preto.