Two rice MADS domain proteins interact with OsMADS1
Two rice MADS domain proteins interact with OsMADS1
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DOI：
10.1023/a:1026517111843
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发表时间：
2000-11-01
影响因子：
5.1
通讯作者：
Jang, SK
中科院分区：
文献类型：
--
作者：
Lim, J;Moon, YH;Jang, SK
OsMADS1 是控制水稻花发育的 MADS 盒基因。为了更多地了解 OsMADS1 的功能，我们利用酵母双杂交系统寻找与 OsMADS1 相互作用的细胞蛋白。从水稻 cDNA 文库中分离出两种具有 MADS 结构域的新型蛋白，命名为 OsMADS14 和 OsMADS15。 OsMADS14 和-15 与玉米 MADS 盒基因 ZAP1 高度同源，ZAP1 是花同源异型基因 APETALA1 (AP1) 的直系同源物。使用大肠杆菌中表达的 GST 融合 OsMADS1 以及 OsMADS14 和 -15 的体外翻译蛋白进行体外实验，证实了这三个 MADS 结构域蛋白之间的相互作用。我们利用双杂交系统和下拉实验确定了 OsMADS1、-14 和 -15 中的哪些结构域是蛋白质-蛋白质相互作用所必需的。虽然 K 结构域对于蛋白质-蛋白质相互作用至关重要，但 K 结构域之前的区域增强了这种相互作用。有趣的是，OsMADS1 的 C 端区域在酵母和哺乳动物细胞中充当转录激活结构域，而另一方面，OsMADS14 和 -15 的 C 结构域仅表现出非常弱的转录激活功能（如果有的话）。
OsMADS1 is a MADS box gene controlling flower development in rice. In order to learn more about the function of OsMADS1, we searched for cellular proteins interacting with OsMADS1 employing the yeast two-hybrid system. Two novel proteins with MADS domains, which were named OsMADS14 and OsMADS15, were isolated from a rice cDNA library. OsMADS14 and -15 are highly homologous to the maize MADS box gene ZAP1 which is an orthologue of the floral homeotic gene APETALA1 (AP1). Interactions among the three MADS domain proteins were confirmed by in vitro experiments using GST-fused OsMADS1 expressed in Escherichia coli and in vitro translated proteins of OsMADS14 and -15. We determined which domains in OsMADS1, -14, and -15 were required for protein-protein interaction employing the two-hybrid system and pull-down experiments. While the K domain was essential for protein-protein interaction, a region preceded by the K domain augmented this interaction. Interestingly, the C-terminal region of OsMADS1 functioned as a transcriptional activation domain in yeast and mammalian cells, while, on the other hand, the C domains of OsMADS14 and -15 exhibited only very weak transcriptional activator functionality, if any at all.