Krolak J M, McClain D, Snyder S L, Fuchs P, Minton K W
Armed Forces Radiobiology Research Institute, Bethesda, Maryland 20814-5145.
Radiat Res. 1989 May;118(2):330-40.
The effects of ionizing radiation (137Cs) on processing of ribosomal RNA (rRNA) were studied by pulse-labeling HeLa S3 cells with [3H]uridine immediately prior to irradiation. The 45 S rRNA precursor, and its two major daughter species, 28 and 18 S rRNA, were separated by gel electrophoresis and the extent of radiolabel incorporation into each was determined at various times after irradiation. This approach permitted kinetic analysis of processing of the 45 S rRNA which had been predominantly synthesized (radiolabeled) prior to irradiation. Since they both derive from the same 45 S pre-rRNA transcript, 28 and 18 S rRNA are produced with a stoichiometry of 1:1, as observed in control cells in the present studies. However, within 1 h following 10 Gy an altered stoichiometry of 28 S:18 S rRNA was apparent, reaching 1.6:1 by 5-7 h following irradiation. This alteration was also observed following the higher dose of 20 Gy, but not following exposures of 5 Gy or less. The 18 S portion of the 45 S pre-rRNA is transcribed prior to the 28 S portion. Consequently, an increase in the 28 S/18 S ratio can only be due to degradation of the 18 S species during or after processing. This alteration may represent a response to radiation-induced growth arrest, by reducing the number of newly synthesized ribosomes that would otherwise be required for cell propagation.
通过在照射前立即用[³H]尿苷脉冲标记HeLa S3细胞,研究了电离辐射(¹³⁷Cs)对核糖体RNA(rRNA)加工的影响。45S rRNA前体及其两个主要子代产物28S和18S rRNA通过凝胶电泳分离,并在照射后的不同时间测定每个产物中放射性标记掺入的程度。这种方法允许对照射前已主要合成(放射性标记)的45S rRNA的加工进行动力学分析。由于28S和18S rRNA都来自相同的45S前体rRNA转录本,如本研究中在对照细胞中观察到的那样,它们以1:1的化学计量比产生。然而,在10 Gy照射后1小时内,28S:18S rRNA的化学计量比发生了明显变化,在照射后5 - 7小时达到1.6:1。在20 Gy的更高剂量照射后也观察到了这种变化,但在5 Gy或更低剂量照射后未观察到。45S前体rRNA的18S部分在28S部分之前转录。因此,28S/18S比值的增加只能归因于加工过程中或加工后18S产物的降解。这种变化可能代表了对辐射诱导的生长停滞的一种反应,通过减少细胞增殖所需的新合成核糖体的数量。