Purification and crystallization of a putative transcriptional regulator of the benzoate oxidation pathway in Burkholderia xenovorans LB400.
Burkholderia xenovorans LB400 harbours two paralogous copies of the recently discovered benzoate oxidation (box) pathway. While both copies are functional, the paralogues are differentially regulated and flanked by putative transcriptional regulators from distinct families. The putative LysR-type transcriptional regulator (LTTR) adjacent to the megaplasmid-encoded box enzymes, Bxe_C0898, has been produced recombinantly in Escherichia coli and purified to homogeneity. Gel-filtration studies show that Bxe_C0898 is a tetramer in solution, consistent with previously characterized LTTRs. Bxe_C0898 crystallized with four molecules in the asymmetric unit of the P4(3)2(1)2/P4(1)2(1)2 unit cell with a solvent content of 61.19%, as indicated by processing of the X-ray diffraction data. DNA-protection assays are currently under way in order to identify potential operator regions for this LTTR and to define its role in regulation of the box pathway.
Law
Adrienne M
AM
Department of Biochemistry and Microbiology, University of Victoria, PO Box 3055 STN CSC, Victoria, BC V8W 3P6, Canada.
Bains
Jasleen
J
Boulanger
Martin J
MJ
D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification*; D001565:Benzoates / Q000378:metabolism*; D019117:Burkholderia / Q000378:metabolism*; D003001:Cloning, Molecular; D003460:Crystallization; D010084:Oxidation-Reduction; D014157:Transcription Factors / Q000737:chemistry / Q000302:isolation & purification*
D001426:Bacterial Proteins; D001565:Benzoates; D014157:Transcription Factors; C073470:LysR protein, Bacteria