Small-angle X-ray scattering and in silico modeling approaches for the accurate functional annotation of an LysR-type transcriptional regulator

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Toledo, M. A. S. ^[1] ; Santos, C. A. ^[1] ; Mendes, J. S. ^[1] ; Pelloso, A. C. ^[1] ; Beloti, L. L. ^[1] ; Crucello, A. ^[1] ; Favaro, M. T. P. ^[1] ; Santiago, A. S. ^[1] ; Schneider, D. R. S. ^[1] ; Saraiva, A. M. ^{[1, 2]} ; Stach-Machado, D. R. ^[3] ; Souza, A. A. ^[4] ; Trivella, D. B. B. ^[5] ; Aparicio, R. ^[5] ; Tasic, L. ^[6] ; Azzoni, A. R. ^{[1, 7]} ; Souza, A. P. ^{[1, 8]}

Afiliação do(s) autor(es)

^[1] Univ Estadual Campinas, Ctr Biol Mol & Engn Genet, Campinas, SP - Brazil

^[2] Inst Nacl Metrol Qualidade & Tecnol, Rio De Janeiro - Brazil

^[3] Univ Estadual Campinas, Inst Biol, Dept Imunol, Campinas, SP - Brazil

^[4] IAC, Ctr APTA Citros Sylvio Moreira, Cordeiropolis, SP - Brazil

^[5] Univ Estadual Campinas, Inst Quim, Lab Biol Estruct & Cristalog, Campinas, SP - Brazil

^[6] Univ Estadual Campinas, Inst Quim, Lab Quim Biol, Campinas, SP - Brazil

^[7] Univ Sao Paulo, Escola Politecn, Dept Engn Quim, Sao Paulo - Brazil

^[8] Univ Estadual Campinas, Dept Biol Vegetal, Inst Biol, Campinas, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS; v. 1834, n. 3, p. 697-707, MAR 2013.
• Citações Web of Science: 5

Resumo

Xylella fastidiosa is a xylem-limited, Gram-negative phytopathogen responsible for economically relevant crop diseases. Its genome was thus sequenced in an effort to characterize and understand its metabolism and pathogenic mechanisms. However, the assignment of the proper functions to the identified open reading frames (ORFs) of this pathogen was impaired due to a lack of sequence similarity in the databases. In the present work, we used small-angle X-ray scattering and in silica modeling approaches to characterize and assign a function to a predicted LysR-type transcriptional regulator in the X. fastidiosa (XfLysRL) genome. XfLysRL was predicted to be a homologue of BenM, which is a transcriptional regulator involved in the degradation pathway of aromatic compounds. Further functional assays confirmed the structural prediction because we observed that XfLysRL interacts with benzoate and cis,cis-muconic acid (also known as 2E,4E-hexa-2,4-dienedioic acid; hereafter named muconate), both of which are co-factors of BenM. In addition, we showed that the XfLysRL protein is differentially expressed during the different stages of X.fastidiosa biofilm formation and planktonic cell growth, which indicates that its expression responds to a cellular signal that is likely related to the aromatic compound degradation pathway. The assignment of the proper function to a protein is a key step toward understanding the cellular metabolic pathways and pathogenic mechanisms. In the context of X. fastidiosa, the characterization of the predicted ORFs may lead to a better understanding of the cellular pathways that are linked to its bacterial pathogenicity. (C) 2012 Published by Elsevier B.V. (AU)

• Processo FAPESP: 01/07533-7 - Resolução da estrutura tridimensional de proteínas relacionadas à patogenicidade de Xyllela fastidiosa
• Beneficiário: Anete Pereira de Souza
• Linha de fomento: Auxílio à Pesquisa - Regular