Screening and analysis of proteins interacting with OsMADS16 in rice (Oryza sativa L.)
Screening and analysis of proteins interacting with OsMADS16 in rice (Oryza sativa L.)
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水稻 (Oryza sativa L.) 中与 OsMADS16 相互作用的蛋白质的筛选和分析
DOI：
10.1371/journal.pone.0221473
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发表时间：
2019-08-22
期刊：
影响因子：
3.7
通讯作者：
Wu, Weiren
中科院分区：
文献类型：
--
作者：
Kong, Lan;Duan, Yuanlin;Wu, Weiren
OsMADS 16是一个B类花器官识别基因，在水稻雄蕊形成中起着关键作用。到目前为止，除了几个MADS盒蛋白外，对OsMADS 16的相互作用伴侣知之甚少。本研究构建了水稻幼穗（< 5 cm）高质量cDNA文库，并以OsMADS 16为诱饵进行酵母双杂交（Y2 H）筛选。通过Y2 H鉴定了11个与OsMADS 16相互作用的候选蛋白，并通过BiFC和Co-IP实验进行了验证。亚细胞定位结果进一步证实了OsMADS 16与10个候选蛋白在天然水稻细胞中相互作用的可能性。生物信息学分析表明，这些伴侣具有多种分子、细胞和生理功能。其中一些已知或可能与生殖事件有关，如雄蕊原基的起始、分化和发育（OsMADS 2、OsMADS 4和OsCOP 9）和花粉发育（OsbHLH 40和Os 6PGDH）。本研究结果为进一步研究OsMADS 16调控花器官发育和花粉形成的机制提供了重要参考。
OsMADS16, a class B floral organ identity gene, plays a pivotal role in stamen formation in rice. To date, little is known about the interacting partners of OsMADS16 except for several MADS-box proteins. In this study, we constructed a high-quality cDNA library of young panicles (< 5 cm in length) and performed yeast two-hybrid (Y2H) screening using OsMADS16 as bait. Eleven candidate proteins interacting with OsMADS16 were identified by Y2H and validated by BiFC and Co-IP assays. Subcellular localization results further confirmed the possibility of the interactions of OsMADS16 with 10 of the candidate proteins in natural rice cells. Bioinformatics analysis indicated that these partners exerted various molecular, cellular and physiological functions. Some of them were known or likely to be related to reproductive events, such as stamen primordium initiation, differentiation and development (OsMADS2, OsMADS4 and OsCOP9) and pollen development (OsbHLH40 and Os6PGDH). Our results provide an important reference for further research on OsMADS16-mediated regulation mechanism on floral organ development and pollen formation.