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首页> 外文期刊>Journal of Molecular Biology Research >In-vitro Characterization of an L-Kynurenine-Responsive Transcription Regulator of the Oxidative Tryptophan Degradation Pathway in Burkholderia xenovorans
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In-vitro Characterization of an L-Kynurenine-Responsive Transcription Regulator of the Oxidative Tryptophan Degradation Pathway in Burkholderia xenovorans

机译：异色伯克霍尔德氏菌氧化色氨酸降解途径的L-Kureurenine响应转录调节剂的体外表征。

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To study the transcriptional regulation of oxidative tryptophan degradation in Burkholderia , we used comparative genomics that focused on the operon containing the genes annotated as kynA , kynU and kynB . In all sequenced Beta-proteobacteria to-date, including Burkholderia , Ralstonia , Collimonas , and Cupriavidus species, there is a conserved AsnC/Lrp family transcriptional regulator (TR) gene located upstream and in the opposite strand of the operon encoding for the oxidative tryptophan degradation genes. In Burkholderia xenovorans the TR is Bxe_A0736. GST-Bxe_A0736 binds L-kynurenine with greater affinity and specificity than any other amino acid or tryptophan degradation product with a dissociation constant of ~82 ± 11 ?M. DNase I footprinting suggested that Bxe_A0736 protects a set of four degenerate, palindromic sequences within the intergenic region between Bxe_A0735 ( kynB ) and Bxe_A0736. The optimal consensus sequence obtained by analysis for these sites, ATATTCCGAATAT, closely resembles the sequence obtained with a protein binding microarray. Under our fluorescence anisotropy experimental conditions, 1 mM L-kynurenine increased the affinity of Bxe_A0736 for a portion of its promoter region. Our results are consistent with Bxe_A0736 acting as the TR that promotes the transcription of the oxidative tryptophan degradation genes in the presence of L-kynurenine while inhibiting the transcription of its own gene.
机译：为了研究伯克霍尔德菌中氧化色氨酸降解的转录调控，我们使用了比较基因组学，着眼于操纵子包含注释为kynA，kynU和kynB的基因。迄今为止，在所有测序的β-变形杆菌中，包括伯克霍尔德菌，罗氏菌，Collimonas和Cupriavidus菌种，在操纵子的上游和相反链中都有一个保守的AsnC / Lrp家族转录调节因子（TR）基因，编码氧化色氨酸。降解基因。在异种伯克霍尔德氏菌中，TR是Bxe_A0736。与任何其他氨基酸或色氨酸降解产物相比，GST-Bxe_A0736与L-犬尿氨酸的亲和力和特异性更高，其解离常数约为82±11？M。 DNase I足迹表明Bxe_A0736保护Bxe_A0735（kynB）和Bxe_A0736之间的基因间区域内的四个简并回文序列集。通过分析这些位点获得的最佳共有序列ATTATCCGAATAT与通过蛋白质结合微阵列获得的序列非常相似。在我们的荧光各向异性实验条件下，1 mM L-犬尿氨酸增加了Bxe_A0736对部分启动子区域的亲和力。我们的结果与Bxe_A0736充当TR一致，后者在存在L-犬尿氨酸的情况下促进氧化色氨酸降解基因的转录，同时抑制其自身基因的转录。

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