Characterization of an oxidative stress response regulator, homologous to Escherichia coli OxyR, from the phytopathogen Xylella fastidiosa.
The OxyR oxidative stress transcriptional regulator is a DNA-binding protein that belongs to the LysR-type transcriptional regulators (LTTR) family. It has the ability to sense oxidative species inside the cell and to trigger the cell's response, activating the transcription of genes involved in scavenging oxidative species. In the present study, we have overexpressed, purified and characterized the predicted OxyR homologue (orf xf1273) of the phytopathogen Xylella fastidiosa. This bacterium is the causal agent of citrus variegated chlorosis (CVC) disease caused by the 9a5c strain, resulting in economic and social losses. The secondary structure of the recombinant protein was analyzed by circular dichroism. Gel filtration showed that XfoxyR is a dimer in solution. Gel shift assays indicated that it does bind to its own predicted promoter under in vitro conditions. However, considering our control experiment we cannot state that this interaction occurs in vivo. Functional complementation assays indicated that xfoxyR is able to restore the oxidative stress response in an oxyr knockout Escherichia coli strain. These results show that the predicted orfxf1273 codes for a transcriptional regulator, homologous to E. coli OxyR, involved in the oxidative stress response. This may be important for X. fastidiosa to overcome the defense mechanisms of its host during the infection and colonization processes.
Toledo
M A S
MA
Centro de Biologia Molecular e Engenharia Genética-CBMEG, Universidade Estadual de Campinas, Brazil.
Schneider
D R
DR
Azzoni
A R
AR
Favaro
M T P
MT
Pelloso
A C
AC
Santos
C A
CA
Saraiva
A M
AM
Souza
A P
AP
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D002942:Circular Dichroism; D003001:Cloning, Molecular; D024202:Electrophoretic Mobility Shift Assay; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D018384:Oxidative Stress; D010944:Plants / Q000378:metabolism / Q000382:microbiology; D011401:Promoter Regions, Genetic / Q000502:physiology; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D017385:Sequence Homology; D014158:Transcription, Genetic / Q000502:physiology; D044167:Xylella / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D012097:Repressor Proteins; C499299:oxyR protein, E coli