NMR structure of the N-terminal domain of SUMO ligase PIAS1 and its interaction with tumor suppressor p53 and A/T-rich DNA oligomers
NMR structure of the N-terminal domain of SUMO ligase PIAS1 and its interaction with tumor suppressor p53 and A/T-rich DNA oligomers
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DOI：
10.1074/jbc.m403561200
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发表时间：
2004-07-23
影响因子：
4.8
通讯作者：
Shindo, H
中科院分区：
文献类型：
--
作者：
Okubo, S;Hara, F;Shindo, H
已报道蛋白质的皮亚斯（活化STAT的蛋白质抑制剂）家族的成员PIAS 1充当肿瘤抑制因子p53及其自身的E3型SUMO连接酶。也有人提出PIAS 1的N-末端结构域与DNA以及p53相互作用。广泛的生物化学研究已致力于最近了解sumoylations和它的生物学意义，而结构方面的皮亚斯家庭和它的相互作用机制与各种因素是不太为人所知的日期。在这项研究中，SUMO连接酶PIAS 1的N-末端结构域（残基1 - 65）的三维结构由NMR光谱确定。该结构揭示了一个独特的四螺旋束，具有上-下-延伸环-下-上的拓扑结构，其中一部分螺旋-延伸环-螺旋代表SAP（SAF-A/B，Acinus和皮亚斯）基序。因此，该N-末端结构域可以被称为四螺旋SAP结构域。谷胱甘肽S-转移酶下拉试验表明，该结构域具有结合肿瘤抑制因子p53，SUMO化的PIAS 1的靶蛋白的能力，而凝胶迁移率试验表明，它对A/T-丰富的DNA有很强的亲和力。四螺旋SAP结构域与16 bp长的DNA复合的NMR分析表明，四螺旋束的一端是结合位点，并可能适合DNA的小沟。结合PIAS 1作为SUMO连接酶的生物学功能，讨论了其三维结构及其结合的双重性。
A member of the PIAS ( protein inhibitor of activated STAT) family of proteins, PIAS1, have been reported to serve as an E3-type SUMO ligase for tumor suppressor p53 and its own. It also was proposed that the N-terminal domain of PIAS1 interacts with DNA as well as p53. Extensive biochemical studies have been devoted recently to understand sumoylations and its biological implications, whereas the structural aspects of the PIAS family and the mechanism of its interactions with various factors are less well known to date. In this study, the three-dimensional structure of the N-terminal domain ( residues 1 - 65) of SUMO ligase PIAS1 was determined by NMR spectroscopy. The structure revealed a unique four-helix bundle with a topology of an up-down-extended loop-down-up, a part of which the helix-extended loop-helix represented the SAP (SAF-A/B, Acinus, and PIAS) motif. Thus, this N-terminal domain may be referred to as a four-helix SAP domain. The glutathione S-transferase pull-down assay demonstrated that this domain possesses a binding ability to tumor suppressor p53, a target protein for sumoylation by PIAS1, whereas gel mobility assays showed that it has a strong affinity toward A/T-rich DNA. An NMR analysis of the four-helix SAP domain complexed with the 16-bp-long DNA demonstrated that one end of the four-helix bundle is the binding site and may fit into the minor groove of DNA. The three-dimensional structure and its binding duality are discussed in conjunction with the biological functions of PIAS1 as a SUMO ligase.