Comparative Proteomic Analysis of Coregulation of CIPK14 and WHIRLY1/3 Mediated Pale Yellowing of Leaves in Arabidopsis.
Comparative Proteomic Analysis of Coregulation of CIPK14 and WHIRLY1/3 Mediated Pale Yellowing of Leaves in Arabidopsis.
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CIPK14 和 WHIRLY1/3 共调节介导拟南芥叶片淡黄化的比较蛋白质组学分析
DOI：
10.3390/ijms19082231
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发表时间：
2018-07-31
影响因子：
5.6
通讯作者：
Miao Y
中科院分区：
文献类型：
--
作者：
Guan Z;Wang W;Yu X;Lin W;Miao Y
在拟南芥突变系钙调磷酸酶b样相互作用蛋白激酶14 （CIPK14）过表达（oeCIPK14）和双敲除WHIRLY1/WHIRLY3 （why1/3）中观察到叶片斑变淡黄。此外，whely1 （WHY1）蛋白在质体和细胞核之间的相对分布受到CIPK14对WHY1磷酸化的影响。为了阐明CIPK14和WHIRLY1/ whirly3介导的叶片淡黄的协同调控，我们对oeCIPK14杂交系（oeCIPK14-var）、why1/3杂交系（why1/3-var）和野生型（WT）进行了差异蛋白质组学分析。利用基质辅助激光解吸电离飞行时间/飞行时间质谱（MALDI-TOF/TOF-MS）鉴定了67个差异丰富蛋白（DAPs）。与WT相比，这67个蛋白中有34个位于oeCIPK14-var系，33个位于why1/3-var系。5个重叠蛋白在oeCIPK14-var系和why1/3-var系中差异表达：atp依赖性Clp蛋白酶水解亚基相关蛋白3 （ClpR3）、二磷酸核酮糖羧化酶大链（RBCL）、β -淀粉酶3 （BAM3）、核糖体再循环因子（RRF）和二磷酸核酮糖羧化酶小链（RBCS）。生物信息学分析表明，大部分DAPs参与光合作用、防御和抗氧化途径、蛋白质代谢、氨基酸代谢、能量代谢、苹果酸生物合成、脂质代谢和转录。因此，在why1/3-var和oeCIPK14-var品系中，叶绿素含量、光系统光化学效率（PS II） （Fv/Fm）和电子传递速率（ETRs）等光系统参数均有所降低，但非光化学猝灭（NPQ）有所增加。两种突变体对强光都表现出高度敏感性。基于对WHY1 /3-var和oeCIPK14-var细胞系DAPs的注释，我们得出结论：叶绿体中CIPK14磷酸化介导的WHY1缺陷与蛋白质代谢受损有关，导致叶绿体功能障碍。
Pale yellowing of leaf variegation is observed in the mutant Arabidopsis lines Calcineurin B-Like-Interacting Protein Kinase14 (CIPK14) overexpression (oeCIPK14) and double-knockout WHIRLY1/WHIRLY3 (why1/3). Further, the relative distribution of WHIRLY1 (WHY1) protein between plastids and the nucleus is affected by the phosphorylation of WHY1 by CIPK14. To elucidate the coregulation of CIPK14 and WHIRLY1/WHIRLY3-mediated pale yellowing of leaves, a differential proteomic analysis was conducted between the oeCIPK14 variegated (oeCIPK14-var) line, why1/3 variegated (why1/3-var) line, and wild type (WT). More than 800 protein spots were resolved on each gel, and 67 differentially abundant proteins (DAPs) were identified by matrix-assisted laser desorption ionization-time of flight/time of flight mass spectrometry (MALDI-TOF/TOF-MS). Of these 67 proteins, 34 DAPs were in the oeCIPK14-var line and 33 DAPs were in the why1/3-var line compared to the WT. Five overlapping proteins were differentially expressed in both the oeCIPK14-var and why1/3-var lines: ATP-dependent Clp protease proteolytic subunit-related protein 3 (ClpR3), Ribulose bisphosphate carboxylase large chain (RBCL), Beta-amylase 3 (BAM3), Ribosome-recycling factor (RRF), and Ribulose bisphosphate carboxylase small chain (RBCS). Bioinformatics analysis showed that most of the DAPs are involved in photosynthesis, defense and antioxidation pathways, protein metabolism, amino acid metabolism, energy metabolism, malate biosynthesis, lipid metabolism, and transcription. Thus, in the why1/3-var and oeCIPK14-var lines, there was a decrease in the photosystem parameters, including the content of chlorophyll, the photochemical efficiency of photosystem (PS II) (Fv/Fm), and electron transport rates (ETRs), but there was an increase in non-photochemical quenching (NPQ). Both mutants showed high sensitivity to intense light. Based on the annotation of the DAPs from both why1/3-var and oeCIPK14-var lines, we conclude that the CIPK14 phosphorylation-mediated WHY1 deficiency in plastids is related to the impairment of protein metabolism, leading to chloroplast dysfunction.
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影响因子：
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