Interactional similarities and differences in the protein complex of PCNA and DNA replication factor C between rice and Arabidopsis
Interactional similarities and differences in the protein complex of PCNA and DNA replication factor C between rice and Arabidopsis
复制标题
水稻和拟南芥PCNA和DNA复制因子C蛋白复合物相互作用的异同
DOI：
10.1186/s12870-019-1874-z
复制
发表时间：
2019
影响因子：
5.3
通讯作者：
Zhao Jie
中科院分区：
文献类型：
--
作者：
Qian Jie;Chen Yueyue;Xu Yaxing;Zhang Xiufeng;Kang Zhuang;Jiao Jinxia;Zhao Jie
背景：增殖细胞核抗原 (PCNA) 是一种保守的三聚环复合物，通过称为复制因子 C (RFC) 的异源五聚体 AAA+ ATP 酶复合物加载到复制叉上，以维持基因组稳定性。尽管酵母中 PCNA-RFC 复合物的结构已被揭示，但高等植物中 PCNA 的功能以及 PCNA-RFC 复合物的蛋白质-蛋白质相互作用尚不清楚。本研究通过酵母双杂交法和双分子荧光互补技术研究了拟南芥和水稻中介导PCNA和RFC亚基相互作用的关键区域。结果：我们观察到OsPCNA可以与所有OsRFC亚基相互作用，而蛋白质-蛋白质相互作用仅存在于拟南芥RFC2/3/4/5和AtPCNA1/2之间。截短分析表明，拟南芥 RFC2/3/4/5 和水稻 RFC1/2 的 C 端对于结合 PCNA 至关重要，而水稻 RFC3/4/5 介导与 PCNA 相互作用的区域分布在 N 端和 C 端。另一方面，我们发现拟南芥和水稻PCNA的C端和N端对PCNA-PCNA相互作用的贡献相同，并且PCNA的域间连接环（IDCL）结构域和C端对于RFC亚基的相互作用是不可缺少的。结论：这些结果表明拟南芥和水稻PCNA在序列、结构和与其他PCNA单体相互作用的模式上高度保守。然而，拟南芥和水稻 RFC 亚基在结合 PCNA 方面也存在显着差异。总而言之，我们的结果可能有助于揭示植物 RFC-PCNA 复合物的生物学功能。
Background:Proliferating cell nuclear antigen (PCNA), a conserved trimeric ring complex, is loaded onto replication fork through a hetero-pentameric AAA+ ATPase complex termed replication factor C (RFC) to maintain genome stability. Although architectures of PCNA-RFC complex in yeast have been revealed, the functions of PCNA and protein-protein interactions of PCNA-RFC complex in higher plants are not very clear. Here, essential regions mediating interactions between PCNA and RFC subunits in Arabidopsis and rice were investigated via yeast-two-hybrid method and bimolecular fluorescence complementation techniques.Results:We observed that OsPCNA could interact with all OsRFC subunits, while protein-protein interactions only exist between Arabidopsis RFC2/3/4/5 and AtPCNA1/2. The truncated analyses indicated that the C-terminal of Arabidopsis RFC2/3/4/5 and rice RFC1/2 is essential for binding PCNA while the region of rice RFC3/4/5 mediating interaction with PCNA distributed both at the N- and C-terminal. On the other hand, we found that the C- and N-terminal of Arabidopsis and rice PCNA contribute equally to PCNA-PCNA interaction, and the interdomain connecting loop (IDCL) domain and C-terminal of PCNAs are indispensable for interacting RFC subunits.Conclusions:These results indicated that Arabidopsis and rice PCNAs are highly conserved in sequence, structure and pattern of interacting with other PCNA monomer. Nevertheless, there are also significant differences between the Arabidopsis and rice RFC subunits in binding PCNA. Taken together, our results could be helpful for revealing the biological functions of plant RFC-PCNA complex.