Seifert H, Scheurlen M, Northemann W, Heinrich P C
Biochim Biophys Acta. 1979 Aug 29;564(1):55-66. doi: 10.1016/0005-2787(79)90188-6.
70-130 S polyparticles as well as 38 S monoparticles were isolated from rat liver nuclei and analyzed in respect to their RNA components by microgel polyacrylamide electrophoresis in formamide. In addition to the high molecular weight polydisperse hnRNA of polyparticles several low molecular weight RNAs (snRNA) were detected. There are at least six distinct snRNA species in polyparticles. Except for one species, which is missing, 38 S monoparticles showed a similar snRNA pattern. From densitometer tracings of microgels the snRNAs were estimated to represent about 11% of the total polyparticle RNA. The number of nucleotides for the various snRNAs were determined from a plot of relative electrophoretic mobility versus log number of nucleotides. The possibility that the snRNAs are degradation products of the hnRNA was excluded on the basis of the following findings. (1) The snRNA pattern was similar in mono- and polyparticles. (2) Whereas the hnRNA of polyparticles incubated at 37 degrees C was extensively degraded, the snRNA did not show a corresponding increase. (3) After a 30 min pulse with [3H]orotate the hn RNA was readily labeled; none of the snRNAs, however, incorporated radioactivity. The snRNAs were still found after treatment of polyparticles with 2 M NaCl excluding contamination by nucleoplasm.
从大鼠肝细胞核中分离出70 - 130S多聚颗粒以及38S单颗粒,并通过在甲酰胺中的微凝胶聚丙烯酰胺电泳对其RNA成分进行分析。除了多聚颗粒的高分子量多分散核不均一RNA(hnRNA)外,还检测到了几种低分子量RNA(snRNA)。多聚颗粒中至少有六种不同的snRNA种类。除了一种缺失的种类外,38S单颗粒显示出类似的snRNA模式。根据微凝胶的密度计追踪结果,估计snRNA约占多聚颗粒总RNA的11%。通过相对电泳迁移率与核苷酸对数的关系图确定了各种snRNA的核苷酸数量。基于以下发现排除了snRNA是hnRNA降解产物的可能性。(1)单颗粒和多聚颗粒中的snRNA模式相似。(2)虽然在37℃孵育的多聚颗粒的hnRNA被广泛降解,但snRNA没有相应增加。(3)用[³H]乳清酸脉冲30分钟后,hnRNA很容易被标记;然而,没有一种snRNA掺入放射性。在用2M NaCl处理多聚颗粒后仍能发现snRNA,排除了核质污染的可能性。