The type VI secretion system (T6SS) is a widespread and versatile protein secretion system found in most Gram- negative bacteria. Studies of T6SS have mainly focused on its role in virulence toward host cells and inter-bacterial inter-actions, but studies have also shown that T6SS4 in Yersinia pseudotuberculosis participates in the acquisition of zinc ions to alleviate the accumulation of hydroxyl radicals induced by multiple stressors. Here, by comparing the gene expression patterns of wild-type and zntR mutant Y. pseudotubercu-losis cells using RNA-seq analysis, T6SS4 and 17 other bio-logical processes were found to be regulated by ZntR. T6SS4 was positively regulated by ZntR in Y. pseudotuberculosis, and further investigation demonstrated that ZntR regulates T6SS4 by directly binding to its promoter region. T6SS4 ex-pression is regulated by zinc via ZntR, which maintains in-tracellular zinc homeostasis and controls the concentration of reactive oxygen species to prevent bacterial death under oxidative stress. This study provides new insights into the regulation of T6SS4 by a zinc-dependent transcriptional regu-lator, and it provides a foundation for further investigation of the mechanism of zinc transport by T6SS.
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