de la Serna I, Cujec T P, Shi Y, Tyler B M
Department of Plant Pathology, University of California, Davis 96516, USA.
Mol Gen Genet. 2000 Jul;263(6):987-94. doi: 10.1007/pl00008699.
In eukaryotes, the levels of ribosomal proteins are coordinately regulated under varying nutritional conditions and at different developmental stages. Little is known about how ribosomal protein levels are coupled to the levels of rRNA. The formation of a ribonucleoprotein particle composed of 5S rRNA and a ribosomal protein is an early step in ribosome assembly. To investigate how these two ribosomal components are regulated in Neurospora crassa, we cloned the gene encoding the 5S rRNA-binding ribosomal protein (crp-4) and developed a novel system for measuring relative 5S rRNA transcriptional rates in vivo, using a reporter RNA derived from the 40S precursor RNA. The reporter RNA is cleaved from the 5S rRNA in vivo and therefore allows us to distinguish between changes in the 5S rRNA transcription rate and 5S rRNA stability. Using this system, we found that transcription of 5S rRNA is constitutive and is not coordinated with the levels of crp-4 mRNA or with 40S rRNA levels during a carbon upshift or a carbon downshift.
在真核生物中,核糖体蛋白的水平在不同的营养条件下和不同的发育阶段受到协同调控。关于核糖体蛋白水平如何与rRNA水平相耦合,我们所知甚少。由5S rRNA和一种核糖体蛋白组成的核糖核蛋白颗粒的形成是核糖体组装的早期步骤。为了研究在粗糙脉孢菌中这两种核糖体组分是如何被调控的,我们克隆了编码与5S rRNA结合的核糖体蛋白(crp - 4)的基因,并开发了一种新系统,利用源自40S前体RNA的报告RNA来测量体内相对5S rRNA的转录速率。该报告RNA在体内从5S rRNA上切割下来,因此使我们能够区分5S rRNA转录速率的变化和5S rRNA的稳定性。使用这个系统,我们发现5S rRNA的转录是组成型的,并且在碳源上调或碳源下调期间,其与crp - 4 mRNA的水平或40S rRNA的水平不协同。