Kawai Tomoko, Fan Jinshui, Mazan-Mamczarz Krystyna, Gorospe Myriam
Box 12, Laboratory of Cellular and Molecular Biology, National Institute on Aging-IRP, National Institutes of Health, 5600 Nathan Shock Drive, Baltimore, MD 21224-6825, USA.
Mol Cell Biol. 2004 Aug;24(15):6773-87. doi: 10.1128/MCB.24.15.6773-6787.2004.
The stability of mRNAs undergoing translation has long been a controversial question. Here, we systematically investigate links between mRNA turnover and translation during the endoplasmic reticulum (ER) stress response, a process during which protein synthesis is potently regulated. cDNA array-based approaches to assess the stability and translational status of each mRNA were devised. First, ER stress-triggered changes in mRNA stability were studied by comparing differences in steady-state mRNA levels with differences in gene transcription. Second, changes in translational status were monitored by studying ER stress-induced shifts in the relative distribution of each mRNA along sucrose gradients. Together, the array-derived data reveal complex links between mRNA stability and translation, with all regulatory groups represented: both stabilized and destabilized mRNAs were found among translationally induced as well as translationally suppressed mRNA collections. Remarkably, however, the subset of stabilized mRNAs was prominently enriched in translationally suppressed transcripts, suggesting that ER stress was capable of causing the stabilization of mRNAs associated with a global reduction in protein synthesis. The cDNA array-based approach described here can be applied to global analyses of mRNA turnover and translation and can serve to investigate subsets of mRNAs subject to joint posttranscriptional control.
正在进行翻译的mRNA的稳定性长期以来一直是一个有争议的问题。在这里,我们系统地研究了内质网(ER)应激反应过程中mRNA周转与翻译之间的联系,在这个过程中蛋白质合成受到有效调控。我们设计了基于cDNA阵列的方法来评估每个mRNA的稳定性和翻译状态。首先,通过比较稳态mRNA水平的差异与基因转录的差异,研究内质网应激引发的mRNA稳定性变化。其次,通过研究内质网应激诱导的每个mRNA沿蔗糖梯度的相对分布变化,监测翻译状态的变化。综合起来,阵列衍生的数据揭示了mRNA稳定性与翻译之间的复杂联系,所有调控组均有体现:在翻译诱导以及翻译抑制的mRNA集合中都发现了稳定和不稳定的mRNA。然而,值得注意的是,稳定的mRNA子集在翻译抑制的转录本中显著富集,这表明内质网应激能够导致与蛋白质合成整体减少相关的mRNA的稳定。这里描述的基于cDNA阵列的方法可应用于mRNA周转和翻译的全局分析,并可用于研究受联合转录后调控的mRNA子集。