O-fucosylation of CPN20 by SPINDLY Derepresses Abscisic Acid Signaling During Seed Germination and Seedling Development.
O-fucosylation of CPN20 by SPINDLY Derepresses Abscisic Acid Signaling During Seed Germination and Seedling Development.
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SPINDLY 对 CPN20 的 O-岩藻糖基化抑制种子萌发和幼苗发育过程中的脱落酸信号传导
DOI：
10.3389/fpls.2021.724144
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发表时间：
2021
影响因子：
5.6
通讯作者：
Fan LM
中科院分区：
文献类型：
--
作者：
Liang L;Wang Q;Song Z;Wu Y;Liang Q;Wang Q;Yang J;Bi Y;Zhou W;Fan LM
SPINDLY参与植物发育的某些方面。然而，最近发现这种蛋白质作为O-岩藻糖基转移酶的性质。在这项研究中，我们表明SPINDLY（SPY）与CPN 20在酵母双杂交和split-luc试验中相互作用，并且相互作用由阿坝促进。CPN 20是叶绿体定位的辅助分子伴侣，其负调节ABAR介导的阿坝信号传导。通过使用电子转移解离-MS/MS分析，两个O-岩藻糖基化位点，在哺乳动物细胞和拟南芥中异位表达的CPN 20中检测到第116位和第119位苏氨酸。通过体外肽O-岩藻糖基化测定证实两个苏氨酸残基处的O-岩藻糖基化。我们进一步表明，CPN 20积累在叶绿体的间谍突变体，表明SPY负调控CPN 20定位在叶绿体中。体内蛋白质降解试验沿着CPN 20的定位行为表明，CPN 20进入叶绿体是由SPY负调控的。遗传分析表明，阿坝不敏感的spy-3在种子萌发和早期幼苗发育方面的表型被部分抑制的cpn 20突变，这表明CPN 20的行为下游SPY在这个阿坝信号通路，并可能存在其他途径与CPN 20平行。总的来说，上述数据支持这样的观点，即SPY对CPN 20的O-岩藻糖基化微调拟南芥中的阿坝信号。
SPINDLY is involved in some aspects of plant development. However, the nature of this protein as an O-fucosyltransferase was recently discovered. In this study, we show that SPINDLY (SPY) interacts with CPN20 in yeast two-hybrid and split-luc assays, and the interaction is promoted by ABA. CPN20 is a chloroplast-localized co-chaperonin that negatively regulates ABAR-mediated ABA signaling. By using Electron Transfer Dissociation-MS/MS analysis, two O-fucosylation sites, e.g., 116th and 119th threonines, were detected in ectopically expressed CPN20 in mammalian cells and in Arabidopsis. The O-fucosylation at both threonine residues was confirmed by in vitro peptide O-fucosylation assay. We further show that CPN20 accumulates in the chloroplast of spy mutants, suggesting that SPY negatively regulates CPN20 localization in the chloroplast. In vivo protein degradation assay along with CPN20 localization behavior suggest that import of CPN20 into the chloroplast is negatively regulated by SPY. Genetic analysis shows that ABA insensitive phenotypes of spy-3 in terms of seed germination and early seedling development are partially suppressed by the cpn20 mutation, suggesting that CPN20 acts downstream of SPY in this ABA signaling pathway and that there may exist other pathways in parallel with CPN20. Collectively, the above data support the notion that the O-fucosylation of CPN20 by SPY fine-tunes ABA signaling in Arabidopsis.
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DOI：
10.1016/j.biocel.2008.10.001
发表时间：
2009-05
影响因子：
4
作者：
Luther, Kelvin B.;Haltiwanger, Robert S.
通讯作者：
Haltiwanger, Robert S.
DOI：
10.1083/jcb.201501101
发表时间：
2015-03-30
期刊：
The Journal of cell biology
影响因子：
--
作者：
Bond MR;Hanover JA
通讯作者：
Hanover JA
DOI：
10.1111/j.1365-313x.2009.03839.x
发表时间：
2009-06
期刊：
The Plant journal : for cell and molecular biology
影响因子：
--
作者：
Cui H;Benfey PN
通讯作者：
Benfey PN
影响因子：
10.5
作者：
Lee, SC;Cheng, H;Peng, JR
通讯作者：
Peng, JR
影响因子：
2.9
作者：
Cui, Hongchang
通讯作者：
Cui, Hongchang