Comparative analysis of activator-Esigma54 complexes formed with nucleotide-metal fluoride analogues.
Comparative analysis of activator-Esigma54 complexes formed with nucleotide-metal fluoride analogues.
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DOI：
10.1093/nar/gkp541
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发表时间：
2009-08
影响因子：
14.9
通讯作者：
Buck M
中科院分区：
文献类型：
--
作者：
Burrows PC;Joly N;Nixon BT;Buck M
含有主要变体σ54因子的细菌RNA聚合酶（RNAP）在特异性激活蛋白水解ATP的反应中形成开放启动子复合物。在这里，我们使用代表ATP基态和过渡态的核苷酸-金属氟化物（BeF和AlF）类似物探测σ54-RNAP（Eσ54）和与大肠杆菌激活蛋白PspF的各种细胞活性（AAA+）结构域相关的ATP酶之间的结合相互作用，这些类似物允许捕获复合物（否则与ATP的结合过于短暂）。我们发现，组织和功能的ADP-BeF-和ADP-AlF-依赖的复合物大大重叠。我们的数据支持激活途径，其中激活剂与Eσ54封闭复合物的初始ATP依赖性结合导致Eσ54相对于转录起始位点的重组。然而，当线性双链DNA存在于初始闭合复合物中时，激活剂和Eσ54闭合复合物之间的核苷酸依赖性结合相互作用本身不足以形成开放启动子复合物。
Bacterial RNA polymerase (RNAP) containing the major variant σ54 factor forms open promoter complexes in a reaction in which specialized activator proteins hydrolyse ATP. Here we probe binding interactions between σ54-RNAP (Eσ54) and the ATPases associated with various cellular activities (AAA+) domain of the Escherichia coli activator protein, PspF, using nucleotide-metal fluoride (BeF and AlF) analogues representing ground and transition states of ATP, which allow complexes (that are otherwise too transient with ATP) to be captured. We show that the organization and functionality of the ADP–BeF- and ADP–AlF-dependent complexes greatly overlap. Our data support an activation pathway in which the initial ATP-dependent binding of the activator to the Eσ54 closed complex results in the re-organization of Eσ54 with respect to the transcription start-site. However, the nucleotide-dependent binding interactions between the activator and the Eσ54 closed complex are in themselves insufficient for forming open promoter complexes when linear double-stranded DNA is present in the initial closed complex.
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影响因子：
16
作者：
Bose, Daniel;Pape, Tillmann;Burrows, Patricia C.;Rappas, Mathieu;Wigneshweraraj, Siva R.;Buck, Martin;Zhang, Xiaodong
通讯作者：
Zhang, Xiaodong
影响因子：
3.6
作者：
Bordes, P;Wigneshweraraj, SR;Buck, M
通讯作者：
Buck, M
影响因子：
5.6
作者：
Burrows, Patricia C.;Wigneshweraraj, Siva R.;Buck, Martin
通讯作者：
Buck, Martin
影响因子：
56.9
作者：
Kim, TK;Ebright, RH;Reinberg, D
通讯作者：
Reinberg, D
DOI：
10.1038/nsb972
发表时间：
2003-10-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
DeLaBarre, B;Brunger, AT
通讯作者：
Brunger, AT