Hofmann G E, Hand S C
Department of EPO Biology, University of Colorado, Boulder 80309-0334.
J Exp Biol. 1992 Mar;164:103-16. doi: 10.1242/jeb.164.1.103.
In response to environmental anoxia, embryos of the brine shrimp Artemia franciscana enter a dormant state during which energy metabolism and development are arrested. The intracellular acidification that correlates with this transition into anaerobic dormancy has been linked to the inhibition of protein synthesis in quiescent embryos. In this study, we have addressed the level of control at which a mechanism mediated by intracellular pH might operate to arrest protein synthesis. Two independent lines of evidence suggest that there is an element of translational control when protein synthesis is arrested in dormant embryos. First, as determined by in vitro translation techniques, there were no significant quantitative differences in mRNA pools in dormant as compared to actively developing embryos. In addition, fluorography of the translation products showed that there are no large qualitative changes in mRNA species when embryos become dormant. These data suggest that there was no net degradation of mRNA pools in dormant embryos and that protein synthesis may therefore be controlled more strongly at translation than at transcription. Second, polysome profile studies showed that dormant embryos possess reduced levels of polysomes relative to those found in cells or active embryos. The disaggregation of polysomes is an indication that the initiation step in protein synthesis is disrupted and is further evidence that the mechanism involved in protein synthesis arrest in dormant Artemia involves translational control.
为应对环境缺氧,卤虫(Artemia franciscana)胚胎进入休眠状态,在此期间能量代谢和发育停止。与向厌氧休眠转变相关的细胞内酸化与静止胚胎中蛋白质合成的抑制有关。在本研究中,我们探讨了细胞内pH介导的机制可能在哪个控制水平上发挥作用以阻止蛋白质合成。两条独立的证据表明,当休眠胚胎中蛋白质合成停止时,存在翻译控制的因素。首先,通过体外翻译技术确定,与活跃发育的胚胎相比,休眠胚胎的mRNA库在数量上没有显著差异。此外,翻译产物的荧光自显影显示,当胚胎进入休眠状态时,mRNA种类没有大的质的变化。这些数据表明,休眠胚胎中mRNA库没有净降解,因此蛋白质合成可能在翻译水平比转录水平受到更强的控制。其次,多核糖体图谱研究表明,相对于细胞或活跃胚胎中的多核糖体,休眠胚胎的多核糖体水平降低。多核糖体的解聚表明蛋白质合成的起始步骤受到破坏,这进一步证明,卤虫休眠时蛋白质合成停止所涉及的机制涉及翻译控制。