THE SOLUTION STRUCTURE OF THE RING FINGER DOMAIN FROM THE ACUTE PROMYELOCYTIC LEUKEMIA PROTO-ONCOPROTEIN PML
THE SOLUTION STRUCTURE OF THE RING FINGER DOMAIN FROM THE ACUTE PROMYELOCYTIC LEUKEMIA PROTO-ONCOPROTEIN PML
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DOI：
10.1002/j.1460-2075.1995.tb07139.x
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发表时间：
1995-04-03
期刊：
影响因子：
11.4
通讯作者：
FREEMONT, PS
中科院分区：
文献类型：
--
作者：
BORDEN, KLB;BODDY, MN;FREEMONT, PS
急性早幼粒细胞白血病（APL）归因于染色体易位事件，其导致包含PML蛋白和视黄酸受体α的融合蛋白。PML通常是核多蛋白复合物（称为ND 10、Kr体、核体、PML致癌结构域或POD）的组分，其在APL疾病状态下被破坏。PML含有许多特征性基序，包括称为RING或C3 HC 4指的Zn 2+结合结构域。在这里，我们描述了PML环指的溶液结构，如通过H-1 NMR方法在生理pH下用r.m.s.所有原子的骨架原子偏差为0.88和1.39埃。包括CD和光谱学在内的其他生物物理学研究表明，PML RING指需要Zn 2+进行自主折叠，并且半胱氨酸用于金属连接。与以前解决的马疱疹病毒IE110环指的结构的比较，显示出显着的差异，表明环基序是结构多样性的。通过在瞬时转染的NIH 3 T3细胞上使用定点诱变和免疫荧光，在体内测试RING结构域在PML核体形成中的作用。独立突变的两对半胱氨酸在每个锌+结合位点防止PML核体的形成，这表明一个完全折叠的环域是必要的，这个过程。这些结果表明PML RING结构域可能参与蛋白质-蛋白质相互作用，这一特征可能与其他RING指结构域相同。
Acute prormyelocytic leukaemia (APL) has been ascribed to a chromosomal translocation event which results in a fusion protein comprising the PML protein and the retinoic acid receptor cc. PML is normally a component of a nuclear multiprotein complex (termed ND10, Kr bodies, nuclear bodies, PML oncogenic domains or PODs) which is disrupted in the APL disease state. PML contains a number of characterized motifs including a Zn2+ binding domain called the RING or C3HC4 finger. Here we describe the solution structure of the PML RING finger as solved by H-1 NMR methods at physiological pH with r.m.s. deviations for backbone atoms of 0.88 and 1.39 Angstrom for all atoms. Additional biophysical studies including CD and optical spectroscopy, show that the PML RING finger requires Zn2+ for autonomous folding and that cysteines are used in metal ligation. A comparison of the structure with the previously solved equine herpes virus IE110 RING finger, shows significant differences suggesting that the RING motif is structurally diverse. The role of the RING domain in PML nuclear body formation was tested in vivo, by using site-directed mutagenesis and immunofluorescence on transiently transfected NIH 3T3 cells. Independently mutating two pairs of cysteines in each of the Zn2+ binding sites prevents PML nuclear body formation, suggesting that a fully folded RING domain is necessary for this process. These results suggest that the PML RING domain is probably involved in protein-protein interactions, a feature which may be common to other RING finger domains.