PALE CRESS binds to plastid RNAs and facilitates the biogenesis of the 50S ribosomal subunit
PALE CRESS binds to plastid RNAs and facilitates the biogenesis of the 50S ribosomal subunit
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DOI：
10.1111/tpj.13662
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发表时间：
2017-11-01
期刊：
影响因子：
7.2
通讯作者：
Manavski, Nikolay
中科院分区：
文献类型：
--
作者：
Meurer, Joerg;Schmid, Lisa-Marie;Manavski, Nikolay
植物特异性PALE CRESS（PAC）蛋白以前已被证明是光合自养生长所必需的。在这里，我们进一步研究PAC蛋白的分子功能。PAC定位于质体类核并形成大的蛋白质和含RNA的巨道尔顿复合物。它与叶绿体RNA的特定子集共免疫沉淀，包括psbK-psbI，ndhF，ndhD和23 S核糖体RNA（rRNA），如RNA免疫沉淀结合高通量RNA测序（RIP-seq）分析所示。此外，它与未成熟的50 S核糖体颗粒共迁移，并特异性结合23 S rRNA体外。这与pac中23 S rRNA水平的严重降低相一致，导致翻译缺陷和质体转录物模式和丰度的相关改变，类似于用翻译抑制剂林可霉素处理的植物。因此，我们的结论是质体核糖体的缺陷导致了pac的表型。此外，PAC水平在相应的突变体的情况下或减少诱导的23 S rRNA的结构变化，证明了体内RNA结构探测。PAC是一种定位于质体类核中的RNA结合蛋白，在体内与23 SrRNA、psbK-psbI、ndhF和ndhD mRNA结合，形成高分子量复合物。PAC对质体核糖体生物发生的显著影响严重影响翻译、叶片着色、植物生长并导致幼苗死亡。
The plant-specific PALE CRESS (PAC) protein has previously been shown to be essential for photoautotrophic growth. Here we further investigated the molecular function of the PAC protein. PAC localizes to plastid nucleoids and forms large proteinaceous and RNA-containing megadalton complexes. It co-immunoprecipitates with a specific subset of chloroplast RNAs including psbK-psbI, ndhF, ndhD, and 23S ribosomal RNA (rRNA), as demonstrated by RNA immunoprecipitation in combination with high throughput RNA sequencing (RIP-seq) analyses. Furthermore, it co-migrates with premature 50S ribosomal particles and specifically binds to 23S rRNA invitro. This coincides with severely reduced levels of 23S rRNA in pac leading to translational deficiencies and related alterations of plastid transcript patterns and abundance similar to plants treated with the translation inhibitor lincomycin. Thus, we conclude that deficiency in plastid ribosomes accounts for the pac phenotype. Moreover, the absence or reduction of PAC levels in the corresponding mutants induces structural changes of the 23S rRNA, as demonstrated by invivo RNA structure probing. Our results indicate that PAC binds to the 23S rRNA to promote the biogenesis of the 50S subunit.Significance Statement PAC is an RNA-binding protein localized in plastid nucleoids that is associated invivo with the 23S rRNA as well as with psbK-psbI, ndhF, and ndhD mRNAs, and forms high molecular weight complexes. The pronounced impact of PAC on the biogenesis of plastid ribosomes severely influences translation, leaf coloration, plant growth and leads to seedling lethality.