Department of Microbiology and Immunology, Brody School of Medicine, East Carolina University, Greenville, North Carolina, USA.
J Bacteriol. 2012 Feb;194(3):561-6. doi: 10.1128/JB.05296-11. Epub 2011 Nov 18.
MntH is the only high-affinity manganese transporter identified in Brucella. A previous study showed that MntH is required for the wild-type virulence of Brucella abortus 2308 in mice (Anderson ES, et al., Infect. Immun. 77:3466-3474, 2009) and indicated that the mntH gene is regulated in a manganese-responsive manner in this strain by a Mur homolog. In the study presented here, the transcriptional start site for mntH in B. abortus 2308 was determined by primer extension analysis. Specific interactions between Mur and the mntH promoter region were demonstrated in an electrophoretic mobility shift assay (EMSA), and a Mur binding site was identified in the -55 to -24 region of the mntH promoter by DNase I footprint analysis. The specificity of the interaction of Mur with the putative Mur box was further evaluated by EMSA employing oligonucleotides in which the consensus nucleotides in this region were substituted. These studies not only confirm a direct role for Mur in the Mn-responsive regulation of mntH expression in Brucella abortus 2308 but also identify the cis-acting elements upstream of mntH that are responsible for this regulation.
MntH 是布鲁氏菌中唯一鉴定出的高亲和力锰转运蛋白。先前的研究表明,MntH 是布鲁氏菌 abortus 2308 在小鼠中野生型毒力所必需的(Anderson ES,等,感染。Immun. 77:3466-3474, 2009),并表明该 mntH 基因在该菌株中以锰反应方式受 Mur 同源物调节。在本研究中,通过引物延伸分析确定了 Brucella abortus 2308 中的 mntH 转录起始位点。在电泳迁移率变动分析 (EMSA) 中证明了 Mur 与 mntH 启动子区域之间的特异性相互作用,并且通过 DNase I 足迹分析鉴定了 mntH 启动子的 -55 至 -24 区域中的 Mur 结合位点。通过 EMSA 进一步评估了 Mur 与假定的 Mur 框的相互作用的特异性,该 EMSA 采用了该区域中保守核苷酸被取代的寡核苷酸。这些研究不仅证实了 Mur 在 Brucella abortus 2308 中 Mn 反应性调节 mntH 表达中的直接作用,而且还确定了负责这种调节的 mntH 上游顺式作用元件。