A small molecule directly inhibits the p53 transactivation domain from binding to replication protein A.
A small molecule directly inhibits the p53 transactivation domain from binding to replication protein A.
复制标题
DOI：
10.1093/nar/gks1291
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发表时间：
2013-02-01
影响因子：
14.9
通讯作者：
Oakley GG
中科院分区：
文献类型：
--
作者：
Glanzer JG;Carnes KA;Soto P;Liu S;Parkhurst LJ;Oakley GG
复制蛋白A（RPA）是DNA复制、修复和DNA损伤信号传导所必需的，具有六个ssDNA结合结构域（DBD），包括位于最大亚基RPA 70的N-末端的DBD-F。该结构域作为p53和其他含有两亲性α螺旋结构域的DNA损伤和修复蛋白的结合位点发挥作用。在这里，我们证明了直接结合的ssDNA和p53（p53 TAD 2）的反式激活结构域2 DBD-F，以及DBD-F-定向dsDNA链分离RPA，所有这些都被抑制的阿朴海松酸（FPA）。FPA直接与RPA结合，导致构象转变，如通过淬灭全长RPA中的固有色氨酸荧光所确定的。FPA的结构类似物提供了对抑制所需的化学性质的了解。最后，我们证实具有R41 E和R43 E突变的RPA不能与p53结合，使dsDNA不稳定并通过FPA淬灭色氨酸荧光，这表明蛋白质结合，DNA调节和抑制剂结合都发生在DBD-F上的同一位点内。FPA对p53-RPA相互作用的破坏可能会干扰p53和RPA的调节功能，从而抑制控制细胞周期和维持人类基因组完整性的细胞途径。
Replication protein A (RPA), essential for DNA replication, repair and DNA damage signalling, possesses six ssDNA-binding domains (DBDs), including DBD-F on the N-terminus of the largest subunit, RPA70. This domain functions as a binding site for p53 and other DNA damage and repair proteins that contain amphipathic alpha helical domains. Here, we demonstrate direct binding of both ssDNA and the transactivation domain 2 of p53 (p53TAD2) to DBD-F, as well as DBD-F-directed dsDNA strand separation by RPA, all of which are inhibited by fumaropimaric acid (FPA). FPA binds directly to RPA, resulting in a conformational shift as determined through quenching of intrinsic tryptophan fluorescence in full length RPA. Structural analogues of FPA provide insight on chemical properties that are required for inhibition. Finally, we confirm the inability of RPA possessing R41E and R43E mutations to bind to p53, destabilize dsDNA and quench tryptophan fluorescence by FPA, suggesting that protein binding, DNA modulation and inhibitor binding all occur within the same site on DBD-F. The disruption of p53–RPA interactions by FPA may disturb the regulatory functions of p53 and RPA, thereby inhibiting cellular pathways that control the cell cycle and maintain the integrity of the human genome.
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影响因子：
3.5
作者：
Glanzer, Jason G.;Liu, Shengqin;Oakley, Gregory G.
通讯作者：
Oakley, Gregory G.
影响因子：
14.9
作者：
Daughdrill, GW;Ackerman, J;Lowry, DF
通讯作者：
Lowry, DF
影响因子：
3.4
作者：
Atilgan, AR;Durell, SR;Bahar, I
通讯作者：
Bahar, I
影响因子：
4.4
作者：
Bussi, Giovanni;Donadio, Davide;Parrinello, Michele
通讯作者：
Parrinello, Michele
影响因子：
8
作者：
Candau, R;Scolnick, DM;Berger, SL
通讯作者：
Berger, SL