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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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Author(s):
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]
Total Authors: 7
Affiliation:
[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
Total Affiliations: 3
Document type: Journal article
Source: Protein Expression and Purification; v. 113, p. 72-78, SEP 2015.
Web of Science Citations: 2
Abstract

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)

FAPESP's process: 12/51580-4 - Molecular and functional characterization of proteins potentially related to the phytopatogenicity of biofilm-forming bacteria Xylella fastidiosa
Grantee:Anete Pereira de Souza
Support type: Regular Research Grants