Structural and dynamic studies of DNA recognition by NF-κB p50 RHR homodimer using methyl NMR spectroscopy.
Structural and dynamic studies of DNA recognition by NF-κB p50 RHR homodimer using methyl NMR spectroscopy.
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DOI：
10.1093/nar/gkac535
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发表时间：
2022-07-08
影响因子：
14.9
通讯作者：
Dyson, H. Jane
中科院分区：
文献类型：
--
作者：
Singh, Amrinder;Martinez-Yamout, Maria A.;Wright, Peter E.;Dyson, H. Jane
涉及高能量稀疏分布的构象亚态的蛋白质动力学通常对蛋白质功能至关重要。本研究利用（13C, 1H）特异标记Ile （δ）、Leu和Val甲基的13C弛豫色散实验，描述了转录因子NF-κB p50 Rel同源区（RHR）的同二聚体（p50）2的动力学特性。Free (p50)2在溶液中表现出高动态，表现出μs-ms的弛豫色散，与基态和高能亚态之间的交换一致。这些波动从dna结合环传播到结构域的核心。在κB DNA的存在下，这些运动受到抑制，但DNA复合物的核磁共振光谱显示p50 RHR同型二聚体与某些κB DNA序列结合的多个局部构象。改变κB DNA的长度和序列揭示了促进复合物单结合构象的两个因素：双链中κB位点的长度和识别基序两端鸟嘌呤核苷酸的对称序列。dna结合环的动态性质，以及p50 RHR与某些κB位点的多重结合构象，与p50同型二聚体与不同κB序列的转录活性差异是一致的。Ile 139 CδH3在p50 DBD和p50 RHR二聚体中以及加入DNA后的弛豫色散谱（右）。每个结构中的il139显示在下面的面板中。
Protein dynamics involving higher-energy sparsely populated conformational substates are frequently critical for protein function. This study describes the dynamics of the homodimer (p50)2 of the p50 Rel homology region (RHR) of the transcription factor NF-κB, using 13C relaxation dispersion experiments with specifically (13C, 1H)-labeled methyl groups of Ile (δ), Leu and Val. Free (p50)2 is highly dynamic in solution, showing μs-ms relaxation dispersion consistent with exchange between the ground state and higher energy substates. These fluctuations propagate from the DNA-binding loops through the core of the domain. The motions are damped in the presence of κB DNA, but the NMR spectra of the DNA complexes reveal multiple local conformations of the p50 RHR homodimer bound to certain κB DNA sequences. Varying the length and sequence of κB DNA revealed two factors that promote a single bound conformation for the complex: the length of the κB site in the duplex and a symmetrical sequence of guanine nucleotides at both ends of the recognition motif. The dynamic nature of the DNA-binding loops, together with the multiple bound conformations of p50 RHR with certain κB sites, is consistent with variations in the transcriptional activity of the p50 homodimer with different κB sequences. Relaxation dispersion profiles for Ile 139 CδH3 in p50 DBD and p50 RHR dimer, and after addition of DNA (right). Ile 139 in each structure is shown in the lower panels.
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影响因子：
7.3
作者：
Cheng CS;Feldman KE;Lee J;Verma S;Huang DB;Huynh K;Chang M;Ponomarenko JV;Sun SC;Benedict CA;Ghosh G;Hoffmann A
通讯作者：
Hoffmann A
影响因子：
64.8
作者：
GHOSH, G;VANDUYNE, G;SIGLER, PB
通讯作者：
SIGLER, PB
影响因子：
2.7
作者：
Lundstrom, Patrik;Vallurupalli, Pramodh;Kay, Lewis E.
通讯作者：
Kay, Lewis E.
影响因子：
0.9
作者：
Singh A;Dyson HJ
通讯作者：
Dyson HJ
影响因子：
2.7
作者：
DELAGLIO, F;GRZESIEK, S;BAX, A
通讯作者：
BAX, A