Katinakis P K, Burdon R H
Biochim Biophys Acta. 1981 Mar 26;653(1):39-51. doi: 10.1016/0005-2787(81)90102-7.
Using poly(A)-Sepharose and poly(U)-Sepharose affinity chromatography, various classes of nuclear RNA can be distinguished in Friend leukaemia cells. One of these contains a poly(A) tract (poly(A)+-RNA) and another lacks a poly(A) tract but has an affinity for poly(A)-Sepharose (poly(A)-u+-RNA). The stability of these two particular nuclear RNA classes was examined by using a 'pulse-chase' technique involving D-glucosamine treatment. Nuclear poly(A)-u+-RNA was found to decay as a single component with a half-life of about 12 min. In contrast, nuclear poly(A)+-RNA appears to consist of at least two distinct metabolic components with half-lives of about 22 min and 120 min. Furthermore, poly(A)-u+-RNA is transported from the nuclei much more rapidly than the poly(A)+-RNA. The 'pulse-chase' approach also allowed a quantitative estimate to be made of the conversion of nuclear poly(A)+-RNA and poly(A)-u+-RNA to cytoplasmic poly(A)-RNA and poly(A)-u+-RNA.
利用聚(A)-琼脂糖和聚(U)-琼脂糖亲和层析,可以区分弗瑞德白血病细胞中的各类核RNA。其中一类含有聚(A)序列(聚(A)⁺-RNA),另一类缺乏聚(A)序列,但对聚(A)-琼脂糖有亲和力(聚(A)-u⁺-RNA)。通过使用涉及D-葡萄糖胺处理的“脉冲追踪”技术,研究了这两类特定核RNA的稳定性。发现核聚(A)-u⁺-RNA作为单一成分衰变,半衰期约为12分钟。相比之下,核聚(A)⁺-RNA似乎至少由两个不同的代谢成分组成,半衰期分别约为22分钟和120分钟。此外,聚(A)-u⁺-RNA从细胞核转运到细胞质的速度比聚(A)⁺-RNA快得多。“脉冲追踪”方法还使得能够对核聚(A)⁺-RNA和聚(A)-u⁺-RNA向细胞质聚(A)-RNA和聚(A)-u⁺-RNA的转化进行定量估计。