Scanning iron response regulator binding sites using Dap-seq in the Brucella genome

文献类型： 外文期刊

第一作者： Zhang, Huan

作者： Zhang, Huan;Sun, Tianhao;Ma, Zhongchen;Wang, Yueli;Yang, Ningning;Xu, Mingguo;Deng, Xiaoyu;Li, Honghuan;Yi, Jihai;Wang, Zhen;Chen, Chuangfu;Zhang, Huan;Sun, Tianhao;Cao, Xudong;Ma, Zhongchen;Wang, Yueli;Yang, Ningning;Xu, Mingguo;Deng, Xiaoyu;Li, Honghuan;Yi, Jihai;Wang, Zhen;Zhang, Qian;Chen, Chuangfu;Cao, Xudong;Wang, Yifan;Wang, Benben;Zhang, Qian

作者机构：

期刊名称：PLOS NEGLECTED TROPICAL DISEASES （ 影响因子：3.8； 五年影响因子：4.1 ）

ISSN： 1935-2735

年卷期： 2023 年 17 卷 7 期

页码：

收录情况： SCI

摘要： Iron is an essential element required for all organisms. Iron response regulator (Irr) is a crucial transcriptional regulator and can affect the growth and iron uptake of Brucella. The growth rate of Brucella melitensis M5-90 irr mutant was significantly lower than that of B. melitensis M5-90 under normal or iron-sufficient conditions, however, the growth rate of the B. melitensis M5-90 irr mutant was significantly higher than that of B. melitensis M5-90 under iron-limited conditions. In addition, irr mutation significantly reduced iron uptake under iron-limited conditions. Previous studies suggested that the Irr protein has multiple target genes in the Brucella genome that are involved in iron metabolism. Therefore, in the present study, a Dap-seq approach was used to investigate the other iron metabolism genes that are also regulated by the Irr protein in Brucella. A total of seven genes were identified as target genes for Irr in this study and the expression levels of these seven genes were similar in both the RNA-seq and qRT-PCR results. The electrophoretic mobility shift assay confirmed that six out of the seven genes, namely rirA (BME_RS13665), membrane protein (BME_RS01725), hypothetical protein (BME_RS09560), ftrA (BME_RS14525), cation-transporting P-type ATPase (zntA) (BME_RS10660), and 2Fe-2S binding protein (BME_RS13655), interact with the Irr protein. Furthermore, the iron utilization and growth assay experiments confirmed that rirA was involve in iron metabolism and growth of Brucella. In summary, our results identified six genes regulated by the Irr protein that may participate in iron metabolism, and the rirA was identified as a regulon of Irr and it also plays a role in iron metabolism of Brucella. Collectively, these results provide valuable insights for the exploration of Brucella iron metabolism. Author summaryIron response regulator (Irr) plays a key role in iron metabolism of Brucella. In this study, the Brucella melitensis M5-90 irr mutant appeared significant different of growth and iron utilization rate compared with that of the Brucella melitensis M5-90 strain. However, the specific binding sites of Irr in Brucella are not fully understood. Here, we used Dap-seq to identify the potential target genes of Irr in the genome of Brucella melitensis M5-90. A total of seven genes were identified as target genes for Irr and the expression of levels of these seven genes were identified using qRT-PCR. In addition, Six out of seven genes namely rirA, membrane protein, hypothetical protein, ftrA, zntA and 2Fe-2S binding protein interact with the Irr protein. Furthermore, the mutation of rirA markedly affected the iron utilization and growth rate of Brucella. Overall, these results provide valuable insights for the exploration of Brucella iron metabolism.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序