Specific association of estrogen receptor β with the cell cycle spindle assembly checkpoint protein, MAD2
Specific association of estrogen receptor β with the cell cycle spindle assembly checkpoint protein, MAD2
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DOI：
10.1073/pnas.050580997
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发表时间：
2000-03-14
影响因子：
11.1
通讯作者：
Mendelsohn, ME
中科院分区：
文献类型：
--
作者：
Poelzl, G;Kasai, Y;Mendelsohn, ME
雌激素受体（er）是配体激活的转录因子，调节基因表达和细胞生长。目前已经确定了两种内质网：内质网α和最近发现的内质网β。内质网β的生理功能尚不清楚，但来自血管损伤研究和内质网β敲除小鼠的证据表明，内质网β可能参与细胞增殖的调节。在这里，我们展示了内质网β与细胞周期有丝分裂纺锤体组装检查点蛋白MAD2（有丝分裂抑制缺陷2）之间的直接和特异性相互作用。通过筛选酵母双杂交系统血管内皮细胞库，确定了ER β - mad2相互作用，并通过谷胱甘肽s -转移酶融合蛋白相互作用研究证实了这一相互作用。相比之下，ER α在双杂交系统或蛋白-蛋白相互作用实验中均未与MAD2相互作用。在双杂交体系和谷胱甘肽s -转移酶融合蛋白研究中，ER β铰链区的氨基酸173-208足以介导与MAD2的相互作用。这些数据确定了内质网β和MADZ之间的联系，对细胞周期的调节具有潜在的重要性，并支持内质网β的功能不同于已知的内质网作为转录因子的作用。
Estrogen receptors (ERs) are ligand-activated transcription factors that regulate gene expression and cell growth. Two ERs now have been identified: ER alpha and the more recently discovered ER beta, The physiological function of ER beta remains unclear, but evidence from vascular injury studies and from ER beta knockout mice suggests that ER beta may be involved in the regulation of cellular proliferation. Here we show a direct and specific interaction between ER beta and the cell cycle mitotic spindle assembly checkpoint protein, MAD2 (mitosis arrest-deficient 2). The ER beta-MAD2 interaction was identified by screening of a yeast two-hybrid system vascular endothelial cell library with ER beta and confirmed with glutathione S-transferase-fusion protein interaction studies. In contrast, ER alpha did not interact with MAD2 in either the two-hybrid system or in the protein-protein interaction experiments. Amino acids 173-208 in the hinge region of ER beta were sufficient to mediate the interaction with MAD2 in the two-hybrid system and in glutathione S-transferase-fusion protein studies. These data identify a link between ER beta and MADZ of potential importance to regulation of the cell cycle and support a function of ER beta distinct from the established role of ERs as transcription factors.