Pei R, Calame K
Department of Biological Chemistry, University of California, Los Angeles 90024.
Mol Cell Biol. 1988 Jul;8(7):2860-8. doi: 10.1128/mcb.8.7.2860-2868.1988.
We have developed a simple cell-free system for studying the stability of different mRNAs in vitro. We demonstrate that the threefold greater stability in vivo of truncated c-myc mRNA (lacking exon 1) compared with that of full-length c-myc mRNA is maintained in our in vitro system. Chimeric mRNAs in which the first exon of c-myc was fused to immunoglobulin C alpha heavy chain or glyceraldehyde-3-phosphate dehydrogenase mRNAs were not rapidly degraded, demonstrating that c-myc exon 1 alone is not sufficient to tag mRNAs for rapid degradation. Competition experiments show that full-length c-myc mRNA is specifically recognized by a factor(s) responsible for its rapid degradation. This system will allow further characterization and purification of these factors.
我们开发了一种简单的无细胞系统,用于在体外研究不同mRNA的稳定性。我们证明,在我们的体外系统中,截短的c-myc mRNA(缺少外显子1)在体内的稳定性比全长c-myc mRNA高三倍。c-myc的第一个外显子与免疫球蛋白Cα重链或甘油醛-3-磷酸脱氢酶mRNA融合的嵌合mRNA不会迅速降解,这表明单独的c-myc外显子1不足以标记mRNA进行快速降解。竞争实验表明,全长c-myc mRNA被负责其快速降解的一种或多种因子特异性识别。该系统将允许对这些因子进行进一步的表征和纯化。