Department of Biochemistry and Molecular Biology, University of Miami Miller School of Medicine, Miami, FL 33136, USA.
Nucleic Acids Res. 2018 Nov 16;46(20):11048-11060. doi: 10.1093/nar/gky808.
DEAD-Box proteins (DBPs) constitute a prominent class of RNA remodeling factors that play a role in virtually all aspects of RNA metabolism. To better define their cellular functions, deletions in the genes encoding each of the Escherichia coli DBPs were combined with mutations in genes encoding different Ribonucleases (RNases). Significantly, double-deletion strains lacking Ribonuclease R (RNase R) and either the DeaD or SrmB DBP were found to display growth defects and an enhanced accumulation of ribosomal RNA (rRNA) fragments. As RNase R is known to play a key role in removing rRNA degradation products, these observations initially suggested that these two DBPs could be directly involved in the same process. However, additional investigations indicated that DeaD and SrmB-dependent rRNA breakdown is caused by delays in ribosome assembly that increase the exposure of nascent RNAs to endonucleolytic cleavage. Consistent with this notion, mutations in factors known to be important for ribosome assembly also resulted in enhanced rRNA breakdown. Additionally, significant levels of rRNA breakdown products could be visualized in growing cells even in the absence of assembly defects. These findings reveal a hitherto unappreciated mechanism of rRNA degradation under conditions of both normal and abnormal ribosome assembly.
DEAD-Box 蛋白(DBP)构成了一类重要的 RNA 重塑因子,在 RNA 代谢的几乎所有方面都发挥作用。为了更好地定义它们的细胞功能,将编码大肠杆菌中每个 DBP 的基因缺失与编码不同核糖核酸酶(RNase)的基因中的突变相结合。值得注意的是,缺乏核糖核酸酶 R(RNase R)和 DeaD 或 SrmB DBP 的双缺失菌株显示出生长缺陷和核糖体 RNA(rRNA)片段的积累增强。由于已知 RNase R 在去除 rRNA 降解产物中起着关键作用,这些观察结果最初表明这两个 DBP 可能直接参与同一过程。然而,进一步的研究表明,DeaD 和 SrmB 依赖性 rRNA 断裂是由于核糖体组装的延迟导致新生 RNA 暴露于内切核酸酶切割增加所致。与这一观点一致,已知对核糖体组装很重要的因子的突变也导致 rRNA 断裂增加。此外,即使在没有组装缺陷的情况下,在生长细胞中也可以观察到大量的 rRNA 断裂产物。这些发现揭示了在正常和异常核糖体组装条件下 rRNA 降解的一种以前未被认识的机制。