Law Adrienne M, Bains Jasleen, Boulanger Martin J
Department of Biochemistry and Microbiology, University of Victoria, PO Box 3055 STN CSC, Victoria, BC V8W 3P6, Canada.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Oct 1;65(Pt 10):1001-3. doi: 10.1107/S1744309109032321. Epub 2009 Sep 23.
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.
嗜麦芽窄食单胞菌LB400含有最近发现的苯甲酸氧化（box）途径的两个旁系同源拷贝。虽然这两个拷贝都具有功能，但旁系同源物受到不同的调控，并且两侧是来自不同家族的假定转录调节因子。与大质粒编码的box酶Bxe_C0898相邻的假定赖氨酸R型转录调节因子（LTTR）已在大肠杆菌中重组表达并纯化至同质。凝胶过滤研究表明，Bxe_C0898在溶液中是四聚体，这与先前表征的LTTR一致。如X射线衍射数据处理所示，Bxe_C0898在P4(3)2(1)2/P4(1)2(1)2晶胞的不对称单元中与四个分子结晶，溶剂含量为61.19%。目前正在进行DNA保护试验，以确定该LTTR的潜在操纵子区域，并确定其在box途径调控中的作用。
