Two NCA1 isoforms interact with catalase in a mutually exclusive manner to redundantly regulate its activity in rice
Two NCA1 isoforms interact with catalase in a mutually exclusive manner to redundantly regulate its activity in rice
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两种 NCA1 亚型以相互排斥的方式与过氧化氢酶相互作用，过度调节其在水稻中的活性
DOI：
10.1186/s12870-019-1707-0
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发表时间：
2019-03-18
影响因子：
5.3
通讯作者：
Peng, Xinxiang
中科院分区：
文献类型：
--
作者：
Liu, Jianzhe;Cui, Lili;Peng, Xinxiang
背景NCA1（无过氧化氢酶活性1）最近在拟南芥中被鉴定为一种伴侣蛋白，通过维持CAT折叠来调节过氧化氢酶（CAT）活性。该基因主要存在于高等植物中；有些植物，如拟南芥，只含有一个NCA1基因，而另一些植物，如水稻则含有两个拷贝。目前尚不清楚这两种亚型是否以及如何在这些含有双拷贝的植物物种中发挥调节CAT活性的作用。结果在这项研究中，我们首先注意到NCA1a和NCA1b的时空表达模式在水稻植物中非常相似。随后的 BiFC 和酵母三杂交实验表明，NCA1a 和 NCA1b 都与 CAT 表现出相互排斥的相互作用，而不是同时的相互作用。为了进一步进行功能分析，利用CRISPR/Cas9技术产生了水稻的nca1a和nca1b单突变体或双突变体。在正常和盐度胁迫条件下对这些突变体进行分析发现，与WT相比，einca1aornca1b单突变体在表型和CAT活性方面没有表现出差异，而双突变体始终表现出非常低的CAT活性（约5％）和严重的病变表型。结论这些结果表明NCA1a和NCA1b与CAT表现出互斥的相互作用，以功能冗余的方式调节CAT活性。米饭。
BackgroundNCA1 (NO CATALASE ACTIVITY 1) was recently identified inArabidopsisas a chaperone protein to regulate catalase (CAT) activity through maintaining the folding of CAT. The gene exists mainly in higher plants; some plants, such asArabidopsis, contain only oneNCA1gene, whereas some others such as rice harbor two copies. It is not yet understood whether and how both isoforms have functioned to regulate CAT activity in those two-copy-containing plant species.ResultsIn this study, we first noticed that the spatiotemporal expression patterns ofNCA1aandNCA1bwere very similar in rice plants. Subsequent BiFC and yeast three-hybrid experiments demonstrated that both NCA1a and NCA1b show mutually exclusive, rather than simultaneous, interaction with CAT. For a further functional analysis,nca1aandnca1bsingle mutants or double mutants of rice were generated by CRISPR/Cas9. Analysis on these mutants under both normal and salinity stress conditions found that, as compared with WT, eithernca1aornca1bsingle mutant showed no difference at phenotypes and CAT activities, whereas the double mutants constantly displayed very low CAT activity (about 5%) and serious lesion phenotypes.ConclusionsThese results suggest that NCA1a and NCA1b show mutually exclusive interaction with CAT to regulate CAT activity in a functionally-redundant manner in rice.