Identification and Characterization of the zosA Gene Involved in Copper Uptake in Bacillus subtilis 168
Identification and Characterization of the zosA Gene Involved in Copper Uptake in Bacillus subtilis 168
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枯草芽孢杆菌 168 中参与铜吸收的 zosA 基因的鉴定和表征
DOI：
10.1080/09168451.2015.1107462
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
T.
中科院分区：
文献类型：
--
作者：
Fukuhara;T.;Kobayashi;K.;Kanayama;Y.;Enomoto;S.;Kondo;T.;Tsunekawa;N.;Nemoto;M.;Ogasawara;N.;Inagaki;K.;and Tamura;T.
在Cu ~（2+）限制条件下，dl-青霉胺对枯草芽孢杆菌168的生长有明显的抑制作用。基因芯片和2，602个敲除突变体的筛选结果表明，zosA基因在0.1%dl-青霉胺存在下被去阻遏，且zosA突变体对dl-青霉胺敏感。即使没有螯合剂，ThezosA突变体也延迟了Cu限制下的生长，并且通过使用0.3 mM IPTG和直接插入thezosA基因上游的Pspace启动子的诱导，逆转了对dl-青霉胺的敏感性。此外，该zosA突变体表现出增加的耐受过量的Cu 2+，但没有过量的Zn 2+添加到LB和合成培养基。ZosA蛋白的同源模建表明，该蛋白可以折叠成构成金属转运ATP酶的必需结构域。我们的研究表明zosA是一个参与铜吸收的候选基因。
dl-Penicillamine, a copper-specific metal chelator, remarkably suppressed the growth ofBacillus subtilis168 when added to a synthetic medium under Cu2+limitation. DNA microarray and screening of 2,602 knockout mutants showed that thezosAgene was de-repressed in the presence of 0.1%dl-penicillamine, and that thezosAmutant was sensitive todl-penicillamine medium. ThezosAmutant delayed the growth under Cu-limitation even without the chelator, and the sensitivity todl-penicillamine was reversed by induction using 0.3 mM IPTG and thePspacpromoter inserted directly upstream of thezosAgene. Furthermore, thezosAmutant showed elevated tolerance of excessive Cu2+but not of excessive Zn2+added to LB and synthetic media. Homology modeling of the ZosA protein suggested that the protein can fold itself into essential domains for constituting a metal transporting ATPase. Our study suggests thatzosAis a candidate gene involved in copper uptake.