Hofmann G E, Hand S C
Department of Environmental, Population, and Organismic Biology, University of Colorado, Boulder 80309-0334.
Proc Natl Acad Sci U S A. 1994 Aug 30;91(18):8492-6. doi: 10.1073/pnas.91.18.8492.
Comparing the translational capacities of cell-free systems from aerobically developing embryos of the brine shrimp Artemia franciscana vs. quiescent embryos has revealed a global arrest of protein synthesis. Incorporation rates of [3H]leucine by lysates from 4-h anoxic embryos were 8% of those from aerobic (control) embryos, when assayed at the respective pH values measured for each treatment in vivo. Exposure of embryos to 4 h of aerobic acidosis (elevated CO2 in the presence of oxygen) suppressed protein synthesis to 3% of control values. These latter two experimental treatments promote developmental arrest of Artemia embryos and, concomitantly, cause acute declines in intracellular pH. When lysates from each treatment were assayed over a range of physiologically relevant pH values (pH 6.4-8.0), amino acid incorporation rates in lysates from quiescent embryos were consistently lower than values for the aerobic controls. Acute reversal of pH to alkaline values during the 6-min assays was not sufficient to return the incorporation rates of quiescent lysates to control values. Thus, a stable alteration in translational capacity of quiescent lysates is indicated. Addition of exogenous mRNA did not rescue the suppressed protein synthesis in quiescent lysates, which suggests that the acute blockage of amino acid incorporation is apparently not due to limitation in message. Thus, the results support a role for intracellular pH as an initial signaling event in translational control during quiescence yet, at the same time, indicate that a direct proton effect on the translational machinery is not the sole proximal agent for biosynthetic arrest in this primitive crustacean.
比较卤虫(Artemia franciscana)需氧发育胚胎与静止胚胎的无细胞系统的翻译能力,发现蛋白质合成全面停滞。当在体内每种处理所测得的各自pH值下进行测定时,4小时缺氧胚胎的裂解物对[3H]亮氨酸的掺入率仅为需氧(对照)胚胎的8%。将胚胎暴露于4小时的需氧酸中毒环境(在有氧存在下升高二氧化碳浓度),可将蛋白质合成抑制至对照值的3%。后两种实验处理会促使卤虫胚胎发育停滞,并同时导致细胞内pH值急剧下降。当在一系列生理相关的pH值范围(pH 6.4 - 8.0)内对每种处理的裂解物进行测定时,静止胚胎裂解物中的氨基酸掺入率始终低于需氧对照的值。在6分钟测定期间将pH值急剧逆转至碱性值,并不足以使静止裂解物的掺入率恢复到对照值。因此,表明静止裂解物的翻译能力发生了稳定变化。添加外源mRNA并不能挽救静止裂解物中被抑制的蛋白质合成,这表明氨基酸掺入的急性阻断显然不是由于信使RNA的限制。因此,这些结果支持细胞内pH在静止期间作为翻译控制中的初始信号事件的作用,但同时也表明质子对翻译机制的直接作用不是这种原始甲壳类动物生物合成停滞的唯一近端因素。