White M W, Degnin C, Hill J, Morris D R
Department of Biochemistry, University of Washington, Seattle 98195.
Biochem J. 1990 Jun 15;268(3):657-60. doi: 10.1042/bj2680657.
S-Adenosylmethionine decarboxylase (AdoMetDC) activity was elevated 18.8-fold in Swiss 3T3 fibroblasts which were depleted of cellular polyamines by using the inhibitor difluoromethylornithine (DFMO). Although the cellular level of AdoMetDC mRNA and the half-life of active AdoMetDC protein were also increased (4.3- and 1.5-fold respectively), together they could not account for the magnitude of the increase in AdoMetDC activity. These data suggested that the translation of AdoMetDC mRNA must be increased in the polyamine-depleted cells to account fully for the elevation in activity. The cellular distribution of AdoMetDC mRNA was examined in the polyamine-depleted cells, and it was found almost exclusively associated with large polysomes. In contrast, AdoMetDC mRNA in untreated controls was very heterogeneous, with the proportion associated with monosomes equal to that associated with large polysomes. The shift of the AdoMetDC message into large polysomes occurred within 18 h after addition of DFMO to the cultures and could be reversed by adding exogenous putrescine. The effect of polyamine depletion on AdoMetDC translation was specific, since there was no change in the distribution in polysomes of either actin mRNA or the translationally controlled mRNA encoding ribosomal protein S16 in the DFMO-inhibited cells. Thus the translational efficiency of AdoMetDC mRNA in vivo is regulated either directly or indirectly by the concentration of intracellular polyamines through a mechanism involving translational initiation, which results in a change in the number of ribosomes associated with this mRNA.
使用抑制剂二氟甲基鸟氨酸(DFMO)使瑞士3T3成纤维细胞中的细胞多胺耗竭后,S-腺苷甲硫氨酸脱羧酶(AdoMetDC)的活性升高了18.8倍。尽管AdoMetDC mRNA的细胞水平和活性AdoMetDC蛋白的半衰期也有所增加(分别增加了4.3倍和1.5倍),但它们共同作用仍无法解释AdoMetDC活性增加的幅度。这些数据表明,在多胺耗竭的细胞中,AdoMetDC mRNA的翻译必须增加,才能充分解释活性的升高。研究了多胺耗竭细胞中AdoMetDC mRNA的细胞分布,发现它几乎完全与大的多核糖体相关。相比之下,未处理对照中的AdoMetDC mRNA非常不均一,与单体相关的比例与与大的多核糖体相关的比例相等。在向培养物中添加DFMO后18小时内,AdoMetDC信息转移到了大的多核糖体中,并且通过添加外源性腐胺可以逆转这种转移。多胺耗竭对AdoMetDC翻译的影响是特异性的,因为在DFMO抑制的细胞中,肌动蛋白mRNA或编码核糖体蛋白S16的翻译控制mRNA在多核糖体中的分布没有变化。因此,体内AdoMetDC mRNA的翻译效率通过涉及翻译起始的机制直接或间接受细胞内多胺浓度的调节,这导致与该mRNA相关的核糖体数量发生变化。