Sperling R, Sperling J, Levine A D, Spann P, Stark G R, Kornberg R D
Mol Cell Biol. 1985 Mar;5(3):569-75. doi: 10.1128/mcb.5.3.569-575.1985.
Transcripts of the CAD gene in Syrian hamster cells are as abundant in the nucleus as in the cytoplasm. This was shown by in situ hybridization of whole cells and by solution and blot hybridization of subcellular fractions. Similar results were obtained both for wild-type cells and for a mutant containing amplified CAD genes in which the level of CAD RNA is 150-fold greater. CAD nuclear RNA is indistinguishable from mature mRNA by gel electrophoresis and blot hybridization. Discrete higher-molecular-weight precursors are undetectable, although the persistence of a short length of intervening sequence in the otherwise fully processed RNA is not excluded. CAD RNA is released from nuclei by sonication in physiological conditions in a ribonucleoprotein form that sediments as a broad peak at about 200S in a sucrose gradient. CAD sequences extracted from nuclei by treatment with EDTA and RNase are found in the 30S particles previously described.
叙利亚仓鼠细胞中CAD基因的转录本在细胞核和细胞质中的丰度相同。全细胞原位杂交以及亚细胞组分的溶液杂交和印迹杂交均证明了这一点。野生型细胞和含有扩增CAD基因的突变体(其中CAD RNA水平高150倍)均得到了类似结果。通过凝胶电泳和印迹杂交,CAD核RNA与成熟mRNA无法区分。尽管不排除在其他方面已完全加工的RNA中存在短长度间隔序列,但无法检测到离散的高分子量前体。在生理条件下,通过超声处理,CAD RNA以核糖核蛋白形式从细胞核中释放出来,在蔗糖梯度中以约200S的宽峰形式沉降。用EDTA和RNase处理从细胞核中提取的CAD序列存在于先前描述的30S颗粒中。