Pratt W B, Toft D O
Department of Pharmacology, University of Michigan Medical School, Ann Arbor 48109, USA.
Endocr Rev. 1997 Jun;18(3):306-60. doi: 10.1210/edrv.18.3.0303.
We have provided a historical perspective on a body of steroid receptor research dealing with the structure and physiological significance of the untransformed 9S receptor that has often confused both novice and expert investigators. The frequent controversies and equivocations of earlier studies were due to the fact that the native, hormone-free state of these receptors is a large multiprotein complex that resisted description for many years because of its unstable and dynamic nature. The untransformed 9S state of the steroid and dioxin receptors has provided a unique system for studying the function of the ubiquitous, abundant, and conserved heat shock protein, hsp90. The hormonal control of receptor association with hsp90 provided a method of manipulating the receptor heterocomplex in a manner that was physiologically meaningful. For several steroid receptors, binding to hsp90 was required for the receptor to be in a native hormone-binding state, and for all of the receptors, hormone binding promoted dissociation of the receptor from hsp90 and conversion of the receptor to the DNA-binding state. Although the complexes between tyrosine kinases and hsp90 were discovered earlier, the hormonal regulation or steroid receptor association with hsp90 permitted much more rapid and facile study of hsp90 function. The observations that hsp90 binds to the receptors through their HBDs and that these domains can be fused to structurally different proteins bringing their function under hormonal control provided a powerful linkage between the hormonal regulation of receptor binding to hsp90 and the initial step in steroid hormone action. Because the 9S receptor hsp90 heterocomplexes could be physically stabilized by molybdate, their protein composition could be readily studied, and it became clear that these complexes are multiprotein structures containing a number of unique proteins, such as FKBP51, FKBP52, CyP-40, and p23, that were discovered because of their presence in these structures. Further analysis showed that hsp90 itself exists in a variety of native multiprotein heterocomplexes independent of steroid receptors and other 'substrate' proteins. Cell-free systems can now be used to study the formation of receptor heterocomplexes. As we outlined in the scheme of Fig. 1, the multicomponent receptor-hsp90 heterocomplex assembly system is being reconstituted, and the importance of individual proteins, such as hsp70, p60, and p23, in the assembly process is becoming recognized. It should be noted that our understanding of the mechanism and purpose of steroid receptor heterocomplex assembly is still at an early stage. We can now speculate on the roles of receptor-associated proteins in receptor action, both as individuals and as a group, but their actual functions are still vague or unknown. We can make realistic models about the chaperoning and trafficking of steroid receptors, but we don't yet know how these processes occur, we don't know where chaperoning occurs in the cell (e.g. Is it limited to the cytoplasm? Is it a diffuse process or does chaperoning occur in association with structural elements?), and, with the exception of the requirement for hormone binding, we don't know the extent to which the hsp90-based chaperone system impacts on steroid hormone action. It is not yet clear how far the discovery of this hsp90 heterocomplex assembly system will be extended to the development of a general understanding of protein processing in the cell. Because this assembly system is apparently present in all eukaryotic cells, it probably performs an essential function for many proteins. The bacterial homolog of hsp90 is not an essential protein, but hsp90 is essential in eukaryotes, and recent studies indicate that the development of the cell nucleus from prokaryotic progenitors was accompanied by the duplication of genes for hsp90 and hsp70 (698). (ABSTRACT TRUNCATED)
我们已从历史角度对一系列类固醇受体研究进行了阐述,这些研究涉及未转化的9S受体的结构和生理意义,这常常让新手和专家研究者都感到困惑。早期研究中频繁出现的争议和含糊不清,是因为这些受体的天然、无激素状态是一种大型多蛋白复合物,由于其不稳定和动态的性质,多年来一直难以描述。类固醇和二恶英受体的未转化9S状态为研究普遍存在、丰富且保守的热休克蛋白hsp90的功能提供了一个独特的系统。受体与hsp90结合的激素调控提供了一种以生理上有意义的方式操纵受体异源复合物的方法。对于几种类固醇受体而言,受体处于天然激素结合状态需要与hsp90结合,而对于所有受体来说,激素结合会促进受体与hsp90解离,并使受体转变为DNA结合状态。尽管酪氨酸激酶与hsp90之间的复合物更早被发现,但类固醇受体与hsp90的激素调控使得对hsp90功能的研究更加迅速和便捷。hsp90通过受体的激素结合结构域(HBD)与受体结合,并且这些结构域可以与结构不同的蛋白质融合,使它们的功能受激素控制,这些观察结果在受体与hsp90结合的激素调控和类固醇激素作用的起始步骤之间建立了强有力的联系。由于9S受体-hsp90异源复合物可以被钼酸盐物理稳定,其蛋白质组成能够很容易地被研究,并且很明显这些复合物是包含许多独特蛋白质的多蛋白结构,如FKBP51、FKBP52、CyP - 40和p23,它们因存在于这些结构中而被发现。进一步分析表明,hsp90本身存在于多种天然多蛋白异源复合物中,独立于类固醇受体和其他“底物”蛋白。现在可以使用无细胞系统来研究受体异源复合物的形成。正如我们在图1的方案中所概述的,多组分受体-hsp90异源复合物组装系统正在被重建,并且诸如hsp70、p60和p23等单个蛋白质在组装过程中的重要性正逐渐被认识到。应该指出的是,我们对类固醇受体异源复合物组装的机制和目的的理解仍处于早期阶段。我们现在可以推测受体相关蛋白在受体作用中的作用,无论是作为个体还是作为一个群体,但它们的实际功能仍然模糊不清或未知。我们可以构建关于类固醇受体伴侣作用和运输的现实模型,但我们还不知道这些过程是如何发生的,我们不知道伴侣作用在细胞中的何处发生(例如,它是否仅限于细胞质?它是一个扩散过程还是与结构元件相关联地发生伴侣作用?),并且除了激素结合的要求外,我们不知道基于hsp90的伴侣系统对类固醇激素作用的影响程度。目前尚不清楚这种hsp90异源复合物组装系统的发现将在多大程度上扩展到对细胞中蛋白质加工的全面理解。因为这种组装系统显然存在于所有真核细胞中,它可能对许多蛋白质执行着重要功能。hsp90的细菌同源物不是必需蛋白,但hsp90在真核生物中是必需的,并且最近的研究表明,细胞核从原核祖先发育而来伴随着hsp90和hsp70基因的复制(698)。(摘要截断)