古菌中 NAD-RNA 种类的鉴定和 ADPR-RNA 的脱帽。
Identification of NAD-RNA species and ADPR-RNA decapping in Archaea.
机构信息
Faculty of Biology, Philipps-Universität Marburg, Marburg, Germany.
Institute of Pharmacy and Molecular Biotechnology (IPMB), Heidelberg University, Heidelberg, Germany.
出版信息
Nat Commun. 2023 Nov 21;14(1):7597. doi: 10.1038/s41467-023-43377-x.
NAD is a coenzyme central to metabolism that also serves as a 5'-terminal cap for bacterial and eukaryotic transcripts. Thermal degradation of NAD can generate nicotinamide and ADP-ribose (ADPR). Here, we use LC-MS/MS and NAD captureSeq to detect and identify NAD-RNAs in the thermophilic model archaeon Sulfolobus acidocaldarius and in the halophilic mesophile Haloferax volcanii. None of the four Nudix proteins of S. acidocaldarius catalyze NAD-RNA decapping in vitro, but one of the proteins (Saci_NudT5) promotes ADPR-RNA decapping. NAD-RNAs are converted into ADPR-RNAs, which we detect in S. acidocaldarius total RNA. Deletion of the gene encoding the 5'-3' exonuclease Saci-aCPSF2 leads to a 4.5-fold increase in NAD-RNA levels. We propose that the incorporation of NAD into RNA acts as a degradation marker for Saci-aCPSF2. In contrast, ADPR-RNA is processed by Saci_NudT5 into 5'-p-RNAs, providing another layer of regulation for RNA turnover in archaeal cells.
NAD 是一种代谢过程中的辅酶，同时也是细菌和真核生物转录本的 5'端帽。NAD 的热降解可以产生烟酰胺和 ADP-核糖（ADPR）。在这里，我们使用 LC-MS/MS 和 NAD 捕获测序来检测和鉴定嗜热模型古菌 Sulfolobus acidocaldarius 和嗜盐中温菌 Haloferax volcanii 中的 NAD-RNA。S. acidocaldarius 的四种 Nudix 蛋白都不能在体外催化 NAD-RNA 脱帽，但其中一种蛋白（Saci_NudT5）可以促进 ADPR-RNA 脱帽。NAD-RNA 被转化为 ADPR-RNA，我们在 S. acidocaldarius 总 RNA 中检测到了这种 RNA。编码 5'-3'外切酶 Saci-aCPSF2 的基因缺失会导致 NAD-RNA 水平增加 4.5 倍。我们提出，NAD 掺入 RNA 可作为 Saci-aCPSF2 的降解标记。相比之下，ADPR-RNA 被 Saci_NudT5 加工成 5'-p-RNAs，为古菌细胞中 RNA 周转提供了另一层调控。
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