Plant Cell Development, Humboldt Universität zu Berlin, Philippstr.13, 10115, Berlin, Germany.
Computational Regulatory Genomics, Humboldt-University Berlin/Max Delbrück Centre for Molecular Medicine, 10115, Berlin, Germany.
Plant J. 2024 Sep;119(6):2737-2752. doi: 10.1111/tpj.16945. Epub 2024 Jul 29.
The protein levels of chloroplast photosynthetic genes and genes related to the chloroplast genetic apparatus vary to adapt to different conditions. However, the underlying mechanisms governing these variations remain unclear. The chloroplast intron Maturase K is encoded within the trnK intron and has been suggested to be required for splicing several group IIA introns, including the trnK intron. In this study, we used RNA immunoprecipitation followed by high-throughput sequencing (RIP-Seq) to identify MatK's preference for binding to group IIA intron domains I and VI within target transcripts. Importantly, these domains are crucial for splice site selection, and we discovered alternative 5'-splice sites in three MatK target introns. The resulting alternative trnK lariat structure showed increased accumulation during heat acclimation. The cognate codon of tRNA-K(UUU) is highly enriched in mRNAs encoding ribosomal proteins and a trnK-matK over-expressor exhibited elevated levels of the spliced tRNA-K(UUU). Ribosome profiling analysis of the overexpressor revealed a significant up-shift in the translation of ribosomal proteins compared to photosynthetic genes. Our findings suggest the existence of a novel regulatory mechanism linked to the abundance of tRNA-K(UUU), enabling the differential expression of functional chloroplast gene groups.
叶绿体光合基因和与叶绿体遗传装置相关的基因的蛋白质水平会发生变化以适应不同的条件。然而,调节这些变化的潜在机制尚不清楚。叶绿体内含子 maturase K 编码在 trnK 内含子中,被认为是剪接几个 IIA 类内含子所必需的,包括 trnK 内含子。在这项研究中,我们使用 RNA 免疫沉淀 followed by high-throughput sequencing(RIP-Seq)来鉴定 MatK 对靶转录物中 IIA 类内含子结构域 I 和 VI 的偏好性结合。重要的是,这些结构域对于剪接位点选择至关重要,我们在三个 MatK 靶内含子中发现了替代的 5'-剪接位点。由此产生的替代 trnK 套索结构在热驯化过程中积累增加。tRNA-K(UUU)的同源密码子在编码核糖体蛋白的 mRNAs 中高度富集,并且 trnK-matK 过表达显示出剪接的 tRNA-K(UUU)水平升高。过表达物的核糖体分析揭示了与核糖体蛋白相比,光合作用基因的翻译有明显的上调。我们的发现表明存在一种与 tRNA-K(UUU)丰度相关的新型调节机制,能够使功能性叶绿体基因群的表达产生差异。