Institute of Molecular and Cell Biology, University of Tartu, Riia 23, Tartu 51010, Estonia.
EMBO Rep. 2011 May;12(5):458-62. doi: 10.1038/embor.2011.47. Epub 2011 Apr 1.
Ribosomes are large ribozymes that synthesize all cellular proteins. As protein synthesis is rate-limiting for bacterial growth and ribosomes can comprise a large portion of the cellular mass, elucidation of ribosomal turnover is important to the understanding of cellular physiology. Although ribosomes are widely believed to be stable in growing cells, this has never been rigorously tested, owing to the lack of a suitable experimental system in commonly used bacterial model organisms. Here, we develop an experimental system to directly measure ribosomal stability in Escherichia coli. We show that (i) ribosomes are stable when cells are grown at a constant rate in the exponential phase; (ii) more than half of the ribosomes made during exponential growth are degraded during slowing of culture growth preceding the entry into stationary phase; and (iii) ribosomes are stable for many hours in the stationary phase. Ribosome degradation occurs in growing cultures that contain almost no dead cells and coincides with a reduction of comparable magnitude in the cellular RNA concentration.
核糖体是一种大型核酶,能够合成所有细胞蛋白质。由于蛋白质合成是细菌生长的限速步骤,并且核糖体可以占细胞质量的很大一部分,因此阐明核糖体周转率对于理解细胞生理学非常重要。尽管人们普遍认为核糖体在生长中的细胞中是稳定的,但由于在常用的细菌模式生物中缺乏合适的实验系统,这一点从未得到严格的检验。在这里,我们开发了一种实验系统,可以直接测量大肠杆菌中的核糖体稳定性。我们发现:(i)当细胞在指数生长期以恒定速率生长时,核糖体是稳定的;(ii)在进入静止期之前,培养物生长速度减慢时,超过一半的在指数生长期合成的核糖体被降解;(iii)在静止期,核糖体可以稳定存在数小时。核糖体降解发生在生长中的培养物中,其中几乎没有死细胞,并且与细胞 RNA 浓度相当的减少幅度相吻合。