Structure and regulatory targets of SCO3201, a highly promiscuous TetR-like regulator of Streptomyces coelicolor M145.
Structure and regulatory targets of SCO3201, a highly promiscuous TetR-like regulator of Streptomyces coelicolor M145.
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DOI：
10.1016/j.bbrc.2014.06.003
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发表时间：
2014-07
影响因子：
3.1
通讯作者：
Delin Xu;P. Waack;Qi-Zhong Zhang;S. Werten;W. Hinrichs;M. Virolle
中科院分区：
文献类型：
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作者：
Delin Xu;P. Waack;Qi-Zhong Zhang;S. Werten;W. Hinrichs;M. Virolle
SCO 3201是TetR家族的一个调节子，在天蓝色链霉菌中过表达时，它是形态分化和抗生素产生的强阻遏物。在这里，我们报告了14个新的推定的调节目标，这种调节器usingin vitroformaldehyde交联的鉴定。使用EMSA，证明了纯化的His 6-SCO 3201与5个调节子（SCO 1716、SCO 1950、SCO 3367、SCO 4009和SCO 5046）的启动子区、推定的组氨酸磷酸酶（SCO 1809）、乙酰转移酶（SCO 0988）和聚酮合酶RedX（SCO 5878）的直接结合。逆转录分析表明，转录调控因子SCO 1950，SCO 4009，SCO 5046，以及SCO 0988和RedX的表达下调，SCO 3201过表达，而SCO 1809和SCO 3367的表达上调。通过对负调控基因的启动子区进行比对，得到了一个共有的结合基序。预测的操纵子位点的位置与SCO 3201对7个启动子中的5个的直接抑制作用一致。进一步解析了SCO 3201的2.1 nm晶体结构，为该调节剂的高度混杂性提供了可能的解释，这种高度混杂性可能是SCO 3201对S.天蓝色。
SCO3201, a regulator of the TetR family, is a strong repressor of both morphological differentiation and antibiotic production when overexpressed inStreptomyces coelicolor. Here, we report the identification of 14 novel putative regulatory targets of this regulator usingin vitroformaldehyde cross-linking. Direct binding of purified His6-SCO3201 was demonstrated for the promoter regions of 5 regulators (SCO1716, SCO1950, SCO3367, SCO4009 and SCO5046), a putative histidine phosphatase (SCO1809), an acetyltransferase (SCO0988) and the polyketide synthase RedX (SCO5878), using EMSA. Reverse transcriptional analysis demonstrated that the expression of the transcriptional regulators SCO1950, SCO4009, SCO5046, as well as of SCO0988 and RedX was down regulated, upon SCO3201 overexpression, whereas the expression of SCO1809 and SCO3367 was up regulated. A consensus binding motif was derivedviaalignment of the promoter regions of the genes negatively regulated. The positions of the predicted operator sites were consistent with a direct repressive effect of SCO3201 on 5 out of 7 of these promoters. Furthermore, the 2.1 Å crystal structure of SCO3201 was solved, which provides a possible explanation for the high promiscuity of this regulator that might account for its dramatic effect on the differentiation process ofS. coelicolor.