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Rules of Expansion: an Updated Consensus Operator Site for the CopR-CopY Family of Bacterial Copper Exporter System Repressors.
mSphere ( IF 4.8 ) Pub Date : 2020-05-27 , DOI: 10.1128/msphere.00411-20
Henrik O'Brien 1 , Joseph W Alvin 1 , Sanjay V Menghani 1 , Yamil Sanchez-Rosario 1 , Koenraad Van Doorslaer 1, 2, 3 , Michael D L Johnson 2, 4, 5
Affiliation  

Copper is broadly toxic to bacteria. As such, bacteria have evolved specialized copper export systems (cop operons) often consisting of a DNA-binding/copper-responsive regulator (which can be a repressor or activator), a copper chaperone, and a copper exporter. For those bacteria using DNA-binding copper repressors, few studies have examined the regulation of this operon regarding the operator DNA sequence needed for repressor binding. In Streptococcus pneumoniae (the pneumococcus), CopY is the copper repressor for the cop operon. Previously, homologs of pneumococcal CopY have been characterized to bind a 10-base consensus sequence T/GACANNTGTA known as the cop box. Using this motif, we sought to determine whether genes outside the cop operon are also regulated by the CopY repressor, which was previously shown in Lactococcus lactis. We found that S. pneumoniae CopY did not bind to cop operators upstream of these candidate genes in vitro. During this process, we found that the cop box sequence is necessary but not sufficient for CopY binding. Here, we propose an updated operator sequence for the S. pneumoniae cop operon to be ATTGACAAATGTAGAT binding CopY with a dissociation constant (Kd) of ∼28 nM. We demonstrate strong cross-species interaction between some CopY proteins and CopY operators, suggesting strong evolutionary conservation. Taken together with our binding studies and bioinformatics data, we propose the consensus operator RNYKACANNYGTMRNY for the bacterial CopR-CopY copper repressor homologs.

中文翻译:

扩展规则:细菌铜出口系统抑制器CopR-CopY系列的最新共识操作员站点。

铜对细菌有广泛的毒性。这样,细菌已经进化专门铜出口系统(COP操纵子)通常由DNA结合/铜响应调节器(其可以是一个或阻遏活化剂),铜伴侣,和铜出口的。对于那些使用结合DNA的铜阻遏物的细菌,很少有研究检查该操纵子关于阻遏物结合所需的操纵子DNA序列的调控。在肺炎链球菌(肺炎球菌),拷贝是用于铜阻遏警察操纵子。以前,肺炎球菌CopY的同系物已被表征为结合10个碱基的共有序列T / GACANNTGTA,称为cop box。我们试图利用这种基序来确定cop操纵子也受CopY阻遏物调节,该阻遏物先前已在乳酸乳球菌中显示。我们发现肺炎链球菌CopY在体外不结合这些候选基因上游的cop操纵子。在此过程中,我们发现cop box序列是必需的,但不足以结合CopY。在这里,我们建议为肺炎链球菌cop操作子更新的操作员序列是具有解离常数(K d的ATTGACAAATGTAGAT结合CopY )〜28 nM。我们展示了一些CopY蛋白和CopY运营商之间强大的跨物种相互作用,表明强烈的进化保守性。结合我们的结合研究和生物信息学数据,我们提出了针对细菌CopR-CopY铜阻遏物同源物的共识算子RNYKACANNYGTMRNY。
更新日期:2020-05-27
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