Georgiev O, Mous J, Birnstiel M L
Nucleic Acids Res. 1984 Nov 26;12(22):8539-51. doi: 10.1093/nar/12.22.8539.
Precursors of Xenopus and sea urchin histone mRNAs were synthesized in vitro with the SP6 transcription system, and their maturation and nucleo-cytoplasmic transport was studied by frog oocyte injection. 3' processing is most rapid for homologous histone messenger sequences and does not require either genuine 5' or specific 3' ends of the precursor, but capping of the 5' terminus strongly influences the efficiency of 3' processing. No generation of 5' histone mRNA ends can be detected when precursors containing 5' spacer sequence extensions are injected into the oocyte nucleus. This finding may have some implications for the question whether histone gene transcription could be polycistronic. Using a novel oocyte technique, we have separated nuclei from cytoplasm and have studied the time course of exit of the processed RNA from the oocyte nucleus into the cytoplasm. The results suggest that RNA maturation and nucleo-cytoplasmic transport are not temporally coupled processes.
利用SP6转录系统在体外合成非洲爪蟾和海胆组蛋白mRNA的前体,并通过蛙卵母细胞注射研究其成熟过程及核质转运。对于同源组蛋白信使序列,3'加工最为迅速,且不需要前体真正的5'端或特定的3'端,但5'末端的加帽强烈影响3'加工的效率。当将含有5'间隔序列延伸的前体注射到卵母细胞核中时,未检测到5'组蛋白mRNA末端的产生。这一发现可能对组蛋白基因转录是否可能是多顺反子的问题具有一定启示。使用一种新颖的卵母细胞技术,我们将细胞核与细胞质分离,并研究了加工后的RNA从卵母细胞核进入细胞质的时间进程。结果表明,RNA成熟和核质转运不是时间上偶联的过程。