Hamlett N V, Landale E C, Davis B H, Summers A O
Department of Biology, Swarthmore College, Pennsylvania 19081.
J Bacteriol. 1992 Oct;174(20):6377-85. doi: 10.1128/jb.174.20.6377-6385.1992.
The mercury resistance (mer) operon of the gram-negative transposon Tn21 encodes not only a mercuric reductase and regulatory genes but also two inner membrane proteins (MerT and MerC) and a periplasmic protein (MerP). Although the merT, merP, and merC genes have been implicated in Hg(II) transport, the individual roles of these genes have not been established. We created in vitro precise deletion and frameshift mutations that eliminated each of the genes singly and in combination. Our results show that both merT and merP are required for Hg(II) binding but that merC is not. Both merT and merP are required for full expression of Hg(II) resistance, but loss of merP is less deleterious than loss of merT. Furthermore, mutations eliminating both merT and merP decrease resistance more than the single mutations do. In contrast, mutating merC had no effect on Hg(II) resistance. Both the merT and merP mutations increase the threshold Hg(II) concentration for induction of merA-lacZ transcriptional fusions and cause an increase in the maximal expression level. In contrast, the merC mutation had little effect on the threshold inducing concentration of Hg(II) but decreased the level of expression. Our results show that merT and merP alone are sufficient to specify a mercury transport system. The role of merC remains obscure.
革兰氏阴性转座子Tn21的汞抗性(mer)操纵子不仅编码一种汞还原酶和调控基因,还编码两种内膜蛋白(MerT和MerC)和一种周质蛋白(MerP)。尽管merT、merP和merC基因与Hg(II)转运有关,但这些基因各自的作用尚未明确。我们创建了体外精确缺失和移码突变,单独或组合消除每个基因。我们的结果表明,Hg(II)结合需要merT和merP,但不需要merC。Hg(II)抗性的完全表达需要merT和merP,但merP缺失的有害性小于merT缺失。此外,消除merT和merP的突变比单个突变更能降低抗性。相比之下,突变merC对Hg(II)抗性没有影响。merT和merP突变均提高了merA-lacZ转录融合诱导的阈值Hg(II)浓度,并导致最大表达水平升高。相比之下,merC突变对Hg(II)的阈值诱导浓度影响不大,但降低了表达水平。我们的结果表明,仅merT和merP就足以构成一个汞转运系统。merC的作用仍然不清楚。