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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.)

Characterization of the LysR-type transcriptional regulator YcjZ-like from Xylella fastidiosa overexpressed in Escherichia coli

Texto completo
Autor(es):
Santiago, Andre S. [1] ; Santos, Clelton A. [1] ; Mendes, Juliano S. [1] ; Toledo, Marcelo A. S. [1] ; Beloti, Lilian L. [1] ; Souza, Alessandra A. [2] ; Souza, Anete P. [1, 3]
Número total de Autores: 7
Afiliação do(s) autor(es):
[1] Univ Estadual Campinas, CBMEG, BR-13083875 Campinas, SP - Brazil
[2] Ctr APTA Citros Sylvio Moreira IAC, Cordeiropolis, SP - Brazil
[3] Univ Estadual Campinas, IB, Dept Biol Vegetal, Campinas, SP - Brazil
Número total de Afiliações: 3
Tipo de documento: Artigo Científico
Fonte: Protein Expression and Purification; v. 113, p. 72-78, SEP 2015.
Citações Web of Science: 2
Resumo

The Xylella fastidiosa 9a5c strain is a xylem-limited phytopathogen that is the causal agent of citrus variegated chlorosis (CVC). This bacterium is able to form a biofilm and occlude the xylem vessels of susceptible plants, which leads to significant agricultural and economic losses. Biofilms are associated with bacterial pathogenicity because they are very resistant to antibiotics and other metal-based chemicals that are used in agriculture. The X fastidiosa YcjZ-like (XfYcjZ-like) protein belongs to the LysR-type transcriptional regulator (LTTR) family and is involved in various cellular functions that range from quorum sensing to bacterial survival. In the present study, we report the cloning, expression and purification of XfYcjZ-like, which was overexpressed in Escherichia coil. The secondary folding of the recombinant and purified protein was assessed by circular dichroism, which revealed that XfYcjZ-like contains a typical alpha/beta fold. An initial hydrodynamic characterization showed that XfYcjZ-like is a globular tetramer in solution. In addition, using a polyclonal antibody against XfYcjZ-like, we assessed the expression profile of this protein during the different developmental phases of X. fastidiosa in in vitro cultivated biofilm cells and demonstrated that XfYcjZ-like is upregulated in planktonic cells in response to a copper shock treatment. Finally, the ability of XfYcjZ-like to interact with its own predicted promoter was confirmed in vitro, which is a typical feature of LysR. Taken together, our findings indicated that the XfYcjZ-like protein is involved in both the organization of the architecture and the maturation of the bacterial biofilm and that it is responsive to oxidative stress. (C) 2015 Elsevier Inc. All rights reserved. (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
Processo FAPESP: 12/51580-4 - Caracterização funcional e molecular de proteínas potencialmente relacionadas a fitopatogenicidade da bactéria formadora de biofilme Xylella fastidiosa
Beneficiário:Anete Pereira de Souza
Linha de fomento: Auxílio à Pesquisa - Regular
Processo FAPESP: 11/50268-4 - Caracterização de três fatores de transcrição pertencentes a família LysR de xulella fastidiosa
Beneficiário:André da Silva Santiago
Linha de fomento: Bolsas no Brasil - Doutorado