DNA recognition by a σ(54) transcriptional activator from Aquifex aeolicus.

DNA recognition by a σ(54) transcriptional activator from Aquifex aeolicus.
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DOI:
10.1016/j.jmb.2014.08.009
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发表时间:
2014-10-23
影响因子:
5.6
通讯作者:
Wemmer DE
Wemmer DE
中科院分区:
生物学2区
文献类型:
--
作者:
Vidangos NK;Heideker J;Lyubimov A;Lamers M;Huo Y;Pelton JG;Ton J;Gralla J;Berger J;Wemmer DE

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细菌σ54-聚合酶的转录起始需要转录激活蛋白的作用。激活剂通过DNA结合结构域结合转录起始位点上游的序列特异性。来自激活剂的结构上表征的DNA结合结构域都属于螺旋-转角-螺旋DNA结合蛋白的反转刺激因子(Fis)家族。本文报道了风产液菌(Aquifex aeolicus)中NtrC 4(4DBD)的DNA结合结构域的游离和DNA结合形式的结构。风产液菌是σ54激活剂NtrC家族的成员。两个NtrC 4结合位点被鉴定为lpxC基因上游(-145和-85碱基对),该基因负责脂质A生物合成的第一个关键步骤。这是lpxC表达中σ54调节的第一个实验证据。4DBD在没有DNA的情况下结晶,并且与-145结合位点复合。这些结构和生物化学数据表明,NtrC 4与DNA结合的方式与其同源物Fis相似。相对于Fis,4DBD结合的更大的序列特异性似乎是由比Fis更大数量的有助于亲和力的碱基特异性接触引起的。
Transcription initiation by bacterial σ54-polymerase requires the action of a transcriptional activator protein. Activators bind sequence-specifically upstream of the transcription initiation site via a DNA-binding domain. The structurally characterized DNA-binding domains from activators all belong to the Factor for Inversion Stimulation (Fis) family of helix-turn-helix DNA-binding proteins. We report here structures of the free and DNA-bound forms of the DNA-binding domain of NtrC4 (4DBD) from Aquifex aeolicus, a member of the NtrC family of σ54 activators. Two NtrC4 binding sites were identified upstream (−145 and −85 base pairs) from the start of the lpxC gene, which is responsible for the first committed step in Lipid A biosynthesis. This is the first experimental evidence for σ54 regulation in lpxC expression. 4DBD was crystallized both without DNA and in complex with the −145 binding site. The structures, together with biochemical data, indicate that NtrC4 binds to DNA in a manner that is similar to that of its close homologue, Fis. The greater sequence specificity for the binding of 4DBD relative to Fis seems to arise from a larger number of base specific contacts contributing to affinity than for Fis.
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