Differential Regulation of the Ribosomal Association of mRNA Transcripts in an Arabidopsis Mutant Defective in Jasmonate-Dependent Wound Response.
Differential Regulation of the Ribosomal Association of mRNA Transcripts in an Arabidopsis Mutant Defective in Jasmonate-Dependent Wound Response.
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DOI：
10.3389/fpls.2021.637959
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发表时间：
2021
影响因子：
5.6
通讯作者：
Koo AJ
中科院分区：
文献类型：
--
作者：
Kimberlin A;Holtsclaw RE;Koo AJ
茉莉酰-L-异亮氨酸 (JA-Ile) 是一种强大的氧脂素，负责植物中全基因组转录重编程，从而导致从生长到防御的重大生理转变。双 T-DNA 插入拟南芥突变体 cyp94b1cyp94b3 (b1b3) 在负责氧化 JA-Ile 的细胞色素 p450、CYP94B1 和 CYP94B3 中存在缺陷，其积累的 JA-Ile 水平高出几倍，但表现出抑制 JA-Ile 依赖性的伤口反应——与预期相反。转录组学和蛋白质组学分析表明，虽然与野生型 (WT) 相比，b1b3 对受伤的转录反应基本没有变化，但发现突变体中的许多蛋白质显着减少，这一点通过标记蛋白的免疫印迹分析得到了验证。为了了解这种蛋白质表型及其对 b1b3 表型的假设贡献，对来自 WT 和 b1b3 的受伤莲座叶样品进行了翻译核糖体亲和纯化 RNA 测序分析。超过 1,600 个基因的转录本不会因受伤而大量改变，但在 WT 叶片受伤后，它们与核糖体的关联发生了变化。与之前的观察结果一致，WT 和 b1b3 之间的 mRNA 转录本总数相似；然而，与核糖体相关的转录物池发生了显着变化。最值得注意的是，与 WT 相比，b1b3 中与核糖体池相关的转录本更少，这可能解释了突变体中许多蛋白质的减少。在b1b3中核糖体相关转录本较少的基因中，有与应激反应、特化代谢、蛋白质代谢、核糖体亚基和转录因子相关的基因，与突变体的生化表型一致。这些结果显示了以前未被认识到的翻译水平调控，这些调控受到植物伤口反应过程中 JA 稳态失调的影响。
Jasmonoyl-L-isoleucine (JA-Ile) is a powerful oxylipin responsible for the genome-wide transcriptional reprogramming in plants that results in major physiological shifts from growth to defense. The double T-DNA insertion Arabidopsis mutant, cyp94b1cyp94b3 (b1b3), defective in cytochrome p450s, CYP94B1 and CYP94B3, which are responsible for oxidizing JA-Ile, accumulates several fold higher levels of JA-Ile yet displays dampened JA-Ile–dependent wound responses—the opposite of what is expected. Transcriptomic and proteomic analyses showed that while the transcriptional response to wounding was largely unchanged in b1b3 compared to wild type (WT), many proteins were found to be significantly reduced in the mutant, which was verified by immunoblot analyses of marker proteins. To understand this protein phenotype and their hypothesized contribution to the b1b3 phenotypes, wounded rosette leaf samples from both WT and b1b3 were subject to a translating ribosome affinity purification RNA sequencing analysis. More than 1,600 genes whose transcripts do not change in abundance by wounding changed their association with the ribosomes after wounding in WT leaves. Consistent with previous observations, the total pool of mRNA transcripts was similar between WT and b1b3; however, the ribosome-associated pool of transcripts was changed significantly. Most notably, fewer transcripts were associated with the ribosome pool in b1b3 than in WT, potentially explaining the reduction of many proteins in the mutant. Among those genes with fewer ribosome-associated transcripts in b1b3 were genes relating to stress response, specialized metabolism, protein metabolism, ribosomal subunits, and transcription factors, consistent with the biochemical phenotypes of the mutant. These results show previously unrecognized regulations at the translational level that are affected by misregulation of JA homeostasis during the wound response in plants.
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影响因子：
4.6
作者：
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通讯作者：
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作者：
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