Plant Sciences, Faculty of Biology, Ludwig-Maximilians-University Munich, Großhaderner Street 2-4, 82152 Planegg-Martinsried, Germany.
Plant Physiology, Faculty of Biology, University of Kaiserslautern, Erwin Schrödinger Street, 67653 Kaiserslautern, Germany.
Plant Physiol. 2019 Jan;179(1):248-264. doi: 10.1104/pp.18.00687. Epub 2018 Nov 8.
The chloroplast hosts photosynthesis and a variety of metabolic pathways that are essential for plant viability and acclimation processes. In this study, we show that the sole plastid UMP kinase (PUMPKIN) in Arabidopsis () associates specifically with the introns of the plastid transcripts -UCC, -UAC, and in vivo, as revealed by RNA immunoprecipitation coupled with deep sequencing (RIP-Seq); and that PUMPKIN can bind RNA efficiently in vitro. Analyses of target transcripts showed that PUMPKIN affects their metabolism. Null alleles and knockdowns of were viable but clearly affected in growth, plastid translation, and photosynthetic performance. In mutants, the levels of many plastid transcripts were reduced, while the amounts of others were increased, as revealed by RNA-Seq analysis. PUMPKIN is a homomultimeric, plastid-localized protein that forms in vivo RNA-containing megadalton-sized complexes and catalyzes the ATP-dependent conversion of UMP to UDP in vitro with properties characteristic of known essential eubacterial UMP kinases. A moonlighting function of PUMPKIN combining RNA and pyrimidine metabolism is discussed.
叶绿体是光合作用和多种代谢途径的宿主,这些途径对植物的生存和适应过程至关重要。在这项研究中,我们表明,拟南芥()中唯一的质体 UMP 激酶(PUMPKIN)与质体转录物 -UCC、-UAC 和 - 的内含子特异性结合,这是通过 RNA 免疫沉淀结合深度测序(RIP-Seq)揭示的;并且 PUMPKIN 可以在体外有效地结合 RNA。对靶转录物的分析表明,PUMPKIN 影响它们的代谢。缺失突变体和 的敲低突变体是可行的,但在生长、质体翻译和光合作用性能方面明显受到影响。在 突变体中,许多质体转录物的水平降低,而其他转录物的水平升高,这是通过 RNA-Seq 分析揭示的。PUMPKIN 是一种同源多聚体、质体定位的蛋白质,它在体内形成含有 RNA 的兆道尔顿大小的复合物,并在体外以具有已知必需的真细菌 UMP 激酶特征的 ATP 依赖性方式将 UMP 转化为 UDP。讨论了 PUMPKIN 结合 RNA 和嘧啶代谢的兼职功能。