Unraveling the Complex Interplay of Fis and IHF Through Synthetic Promoter Engineering

Autor(es)

Oliveira Monteiro, Lummy Maria ^[1] ; Sanches-Medeiros, Ananda ^[1] ; Westmann, Caua Antunes ^[1] ; Silva-Rocha, Rafael ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Ribeirao Preto Med Sch FMRP, Ribeirao Preto - Brazil

• Tipo de documento: Artigo Científico
• Fonte: FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY; v. 8, JUN 18 2020.
• Citações Web of Science: 0

Resumo

Bacterial promoters are usually formed by multiplecis-regulatory elements recognized by a plethora of transcriptional factors (TFs). From those, global regulators are key elements since these TFs are responsible for the regulation of hundreds of genes in the bacterial genome. For instance, Fis and IHF are global regulators that play a major role in gene expression control inEscherichia coli, and usually, multiplecis-regulatory elements for these proteins are present at target promoters. Here, we investigated the relationship between the architecture of thecis-regulatory elements for Fis and IHF inE. coli. For this, we analyze 42 synthetic promoter variants harboring consensuscis-elements for Fis and IHF at different distances from the core -35/-10 region and in various numbers and combinations. We first demonstrated that although Fis preferentially recognizes its consensuscis-element, it can also recognize, to some extent, the consensus-binding site for IHF, and the same was true for IHF, which was also able to recognize Fis binding sites. However, changing the arrangement of thecis-elements (i.e., the position or number of sites) can completely abolish the non-specific binding of both TFs. More remarkably, we demonstrated that combiningcis-elements for both TFs could result in Fis and IHF repressed or activated promoters depending on the final architecture of the promoters in an unpredictable way. Taken together, the data presented here demonstrate how small changes in the architecture of bacterial promoters could result in drastic changes in the final regulatory logic of the system, with important implications for the understanding of natural complex promoters in bacteria and their engineering for novel applications. (AU)

• Processo FAPESP: 18/04810-0 - Desconstruindo a complexidade nas redes regulatórias de formação de biofilme em Escherichia coli
• Beneficiário: Ananda Sanches Medeiros
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto
• Processo FAPESP: 12/22921-8 - Abordagens de biologia sintética para decifrar os mecanismos de integração de sinais em promotores bacterianos complexos
• Beneficiário: Rafael Silva Rocha
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores
• Processo FAPESP: 16/05472-6 - Engenharia e prospecção de novos elementos regulatórios em bactéria através de abordagens de biologia sintética
• Beneficiário: Cauã Antunes Westmann
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 16/19179-9 - Desvendando as relações arquitetura/função de promotores bacterianos complexos utilizando abordagens de biologia sintética
• Beneficiário: Lummy Maria Oliveira Monteiro
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 17/50116-6 - Amando o veneno: bases moleculares do metabolismo e toxicidade do conservante de alimentos de ampla utilização, propionato, em Pseudomonas aeruginosa
• Beneficiário: Rafael Silva Rocha
• Modalidade de apoio: Auxílio à Pesquisa - Regular