Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK.
EMBO J. 1982;1(12):1493-8. doi: 10.1002/j.1460-2075.1982.tb01345.x.
Nuclei isolated from both light-grown and dark-grown leaves of Pisum sativum by Percoll density gradient centrifugation incorporate labelled UTP into RNA when supplemented with the other three nucleoside triphosphates. The RNA is heterodisperse, with transcripts up to at least 25S in size. Among these transcripts are sequences hybridizing to cloned DNA probes for wheat rRNA and two abundant chloroplast polypeptides of Pisum, viz. the small subunit of ribulose bisphosphate carboxylase and a polypeptide of the light-harvesting chlorophyll a/b binding complex. Transcription of small subunit and light-harvesting complex sequences is greater (18-fold and 9-fold, respectively) in nuclei from light-grown leaves than in nuclei from dark-grown leaves. Transcription of ribosomal genes, by contrast, is only doubled by growth in the light. Small subunit and light-harvesting complex sequences transcribed in dark-grown nuclei are not degraded in a 120 min chase. These results suggest that the stimulation of accumulation of small subunit and light-harvesting complex mRNAs by exposure of Pisum seedlings to light is mediated by an increase in transcription rather than by a decrease in RNA degradation.
用 Percoll 密度梯度离心法从豌豆的光生长和暗生长叶片中分离出的细胞核,在补充其他三种核苷三磷酸后,将标记的 UTP 掺入 RNA 中。RNA 是异质的,转录物大小至少可达 25S。这些转录物中包括与克隆小麦 rRNA 和豌豆两种丰富的叶绿体多肽(即核酮糖二磷酸羧化酶小亚基和叶绿素 a/b 结合光收集多肽)的 DNA 探针杂交的序列。与暗生长叶片的细胞核相比,光生长叶片的细胞核中小亚基和光收集复合序列的转录更高(分别为 18 倍和 9 倍)。相比之下,核糖体基因的转录仅因在光下生长而增加一倍。在暗生长细胞核中转录的小亚基和光收集复合序列在 120 分钟的追踪实验中不会降解。这些结果表明,豌豆幼苗暴露在光下会刺激小亚基和光收集复合 mRNA 的积累,这是通过转录增加而不是 RNA 降解减少来介导的。