Jankovic Ivana, Brückner Reinhold
Mikrobielle Genetik, Universitat Tubingen, Tubingen, Germany.
J Mol Microbiol Biotechnol. 2007;12(1-2):114-20. doi: 10.1159/000096466.
Sucrose utilization in Staphylococcus xylosus is dependent on two genes, scrA and scrB; encoding a PTS permease and a sucrose phosphate hydrolase, respectively. The genes are encoded on separate loci and are transcribed from two promoters, P(scrA) and P(scrB), both of which are controlled by the repressor ScrR by binding to the operator sequences O(A) and O(B). In the scrA promoter region, a catabolite-responsive element (cre), operator for the global catabolite control protein CcpA, is also present, but its contribution to scrA regulation has not been determined. Using an integrative promoter probe plasmid, the activities of the promoters P(scrA) and P(scrB) were determined under different growth conditions. Both promoters are induced by sucrose and induction is prevented when glucose is also present. Without a functional CcpA, glucose-mediated prevention of induction is lost, clearly demonstrating that CcpA ensures hierarchical sugar utilization with glucose as preferred substrate. Measurements of promoter activities in the absence of a functional ScrR repressor indicated that CcpA also acts upon the operators O(A) and O(B), albeit not as efficiently as on the genuine cre in P(srcA). Besides determining the choice of the carbon source, CcpA has a second effect on sucrose gene expression. When sucrose is the sole carbon source, sucrose catabolism activates carbon catabolite repression and CcpA prevents full induction of the sucrose utilization genes by partially repressing the scrA promoter. Thus, CcpA-dependent regulation serves as a built-in autoregulatory device to restrict sucrose uptake.
木糖葡萄球菌中蔗糖的利用依赖于两个基因,scrA和scrB;它们分别编码一种磷酸转移酶通透酶和一种蔗糖磷酸水解酶。这些基因位于不同的基因座上,由两个启动子P(scrA)和P(scrB)转录,这两个启动子都通过与操纵序列O(A)和O(B)结合而受阻遏物ScrR控制。在scrA启动子区域,还存在一个分解代谢物反应元件(cre),它是全局分解代谢物控制蛋白CcpA的操纵子,但尚未确定其对scrA调控的作用。使用整合型启动子探针质粒,在不同生长条件下测定了启动子P(scrA)和P(scrB)的活性。两个启动子均由蔗糖诱导,当同时存在葡萄糖时诱导作用被阻止。在没有功能性CcpA的情况下,葡萄糖介导的诱导抑制作用丧失,这清楚地表明CcpA确保以葡萄糖为优先底物进行分级糖利用。在没有功能性ScrR阻遏物的情况下对启动子活性的测量表明,CcpA也作用于操纵序列O(A)和O(B),尽管其效率不如作用于P(srcA)中的真正cre。除了决定碳源的选择外,CcpA对蔗糖基因表达还有第二个作用。当蔗糖是唯一碳源时,蔗糖分解代谢激活碳分解代谢物阻遏,CcpA通过部分抑制scrA启动子来阻止蔗糖利用基因的完全诱导。因此,依赖CcpA的调控作为一种内置的自动调节机制来限制蔗糖摄取。