Aspart L, Cooke R, Delseny M
Biochim Biophys Acta. 1979 Aug 29;564(1):43-54. doi: 10.1016/0005-2787(79)90187-4.
The stability of polyadenylic acid and polyadenylated RNA was investigated in young radish (Raphanus sativus) seedlings. We first studied the decay of poly(A) content, using a [3H]poly(U) assay, following a complete block of transcription by cordycepin (200 microgram/ml). Two lifetime classes of polyadenylic acid have been determined in these seedlings: a short-lived component with a half-life of 30 min which represents 60% of poly(A) and a more stable component with varying half-lives of which the majority range from 4-10 h and a few are considerably longer. During this period rRNA was shown to decay linearly, taking about 41 h for half of this RNA to disappear. The life-time of the other moiety of polyadenylated-RNA was analysed by continuous labelling with [3H]uridine. We have been able to demonstrate that a significant part of the mRNA molecules turns over with a half-life similar to that of the more slowly turning-over poly(A). No evidence could be obtained for rapidly turning-over messenger RNA. Thus the rapidly turning over poly(A) could correspond to a poly(A) turn-over independent of the remainder of the sequence. When labelling was very long, an apparent steady-state was reached and we determined the polyadenylated RNA content of seedlings to be 2.2% of whole cell RNA. Finally, these results were compared with those previously obtained in studying early germination of radish embryo axes. In contrast with stored mRNA which is rapidly degraded following imbibition, part of the mRNA present in 22 h old seedlings is stable for several hours.
我们研究了幼嫩萝卜(Raphanus sativus)幼苗中聚腺苷酸和聚腺苷酸化RNA的稳定性。我们首先使用[3H]聚尿苷酸测定法,在虫草素(200微克/毫升)完全阻断转录后,研究了聚(A)含量的衰减情况。在这些幼苗中确定了两类聚腺苷酸寿命:一类半衰期为30分钟的短寿命组分,占聚(A)的60%;另一类更稳定的组分,半衰期各不相同,大多数在4 - 10小时范围内,少数则长得多。在此期间,rRNA呈线性衰减,约41小时后该RNA的一半消失。通过用[3H]尿苷连续标记分析了聚腺苷酸化RNA其他部分的寿命。我们已经能够证明,相当一部分mRNA分子的周转半衰期与周转较慢的聚(A)相似。没有证据表明存在快速周转的信使RNA。因此,快速周转的聚(A)可能对应于与序列其余部分无关的聚(A)周转。当标记时间很长时,达到了明显的稳态,我们测定幼苗中聚腺苷酸化RNA的含量为全细胞RNA的2.2%。最后,将这些结果与之前研究萝卜胚轴早期萌发时获得的结果进行了比较。与吸水后迅速降解的储存mRNA不同,22小时龄幼苗中存在的部分mRNA能稳定存在数小时。