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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Mechanistic insights into c-di-GMP-dependent control of the biofilm regulator FleQ from Pseudomonas aeruginosa

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Autor(es):
Matsuyama, Bruno Y. [1] ; Krasteva, Petya V. [2, 3, 4] ; Baraquet, Claudine [5] ; Harwood, Caroline S. [5] ; Sondermann, Holger [2] ; Navarro, Marcos V. A. S. [1]
Número total de Autores: 6
Afiliação do(s) autor(es):
[1] Univ Sao Paulo, Inst Fis Sao Carlos, Dept Fis & Ciencia Interdisciplinar, BR-13563120 Sao Carlos, SP - Brazil
[2] Cornell Univ, Coll Vet Med, Dept Mol Med, Ithaca, NY 14853 - USA
[3] Inst Pasteur, Dept Struct Biol & Chem, Unite Biol Struct Secret Bacterienne G5, F-75015 Paris - France
[4] Inst Pasteur, CNRS, UMR 3528, F-75015 Paris - France
[5] Univ Washington, Dept Microbiol, Seattle, WA 98195 - USA
Número total de Afiliações: 5
Tipo de documento: Artigo Científico
Fonte: Proceedings of the National Academy of Sciences of the United States of America; v. 113, n. 2, p. E209-E218, JAN 12 2016.
Citações Web of Science: 42
Resumo

Bacterial biofilm formation during chronic infections confers increased fitness, antibiotic tolerance, and cytotoxicity. In many pathogens, the transition from a planktonic lifestyle to collaborative, sessile biofilms represents a regulated process orchestrated by the intracellular second-messenger c-di-GMP. A main effector for c-di-GMP signaling in the opportunistic pathogen Pseudomonas aeruginosa is the transcription regulator FleQ. FleQ is a bacterial enhancer-binding protein (bEBP) with a central AAA+ ATPase sigma(54)-interaction domain, flanked by a C-terminal helix-turn-helix DNA-binding motif and a divergent N-terminal receiver domain. Together with a second ATPase, FleN, FleQ regulates the expression of flagellar and exopolysaccharide biosynthesis genes in response to cellular c-di-GMP. Here we report structural and functional data that reveal an unexpected mode of c-di-GMP recognition that is associated with major conformational rearrangements in FleQ. Crystal structures of FleQ's AAA+ ATPase domain in its apo-state or bound to ADP or ATP-gamma-S show conformations reminiscent of the activated ring-shaped assemblies of other bEBPs. As revealed by the structure of c-di-GMP-complexed FleQ, the second messenger interacts with the AAA+ ATPase domain at a site distinct from the ATP binding pocket. c-di-GMP interaction leads to active site obstruction, hexameric ring destabilization, and discrete quaternary structure transitions. Solution and cell-based studies confirm coupling of the ATPase active site and c-di-GMP binding, as well as the functional significance of crystallographic interprotomer interfaces. Taken together, our data offer unprecedented insight into conserved regulatory mechanisms of gene expression under direct c-di-GMP control via FleQ and FleQ-like bEBPs. (AU)

Processo FAPESP: 11/24168-2 - Caracterização estrutural e funcional de FleQ de Pseudomonas e Xanthomonas: um importante fator de transcrição envolvido na expressão de genes flagelares e formação de biofilme
Beneficiário:Bruno Yasui Matsuyama
Linha de fomento: Bolsas no Brasil - Doutorado Direto
Processo FAPESP: 09/13238-0 - Estudos estruturais e funcionais de proteínas envolvidas em vias de sinalização celular mediadas por c-di-GMP
Beneficiário:Marcos Vicente de Albuquerque Salles Navarro
Linha de fomento: Auxílio à Pesquisa - Apoio a Jovens Pesquisadores