Wersig C, Bindereif A
Max-Planck-Institut fuer Molekulare Genetik, Otto-Warburg-Laboratorium, Berlin, FRG.
Nucleic Acids Res. 1990 Nov 11;18(21):6223-9. doi: 10.1093/nar/18.21.6223.
U4 snRNA is phylogenetically highly conserved and organized in several domains. To determine the function of each of the domains of human U4 snRNA in the multi-step process of snRNP and spliceosome assembly, we used reconstitution procedures in combination with snRNA mutagenesis. The highly conserved 5' terminal domain of U4 snRNA consists of the stem I and stem II regions that have been proposed to base pair with U6 snRNA, and the 5' stem-loop structure. We found that each of these structural elements is essential for spliceosome assembly. However, only the stem II region is required for U4-U6 interaction, and none of these elements for Sm protein binding. In contrast, the 3' terminal domain of U4 snRNA containing the Sm binding site is dispensable for both U4-U6 interaction and spliceosome assembly. Our results support an organization of the U4 snRNP into multiple functional domains, each of which acts at distinct stages of snRNP and spliceosome assembly.
U4小核仁RNA在系统发育上高度保守，并由几个结构域组成。为了确定人U4小核仁RNA的每个结构域在小核核糖核蛋白（snRNP）和剪接体组装的多步骤过程中的功能，我们将重组程序与小核仁RNA诱变相结合。U4小核仁RNA高度保守的5'末端结构域由已被提出与U6小核仁RNA碱基配对的茎I和茎II区域以及5'茎环结构组成。我们发现这些结构元件中的每一个对于剪接体组装都是必不可少的。然而，只有茎II区域是U4与U6相互作用所必需的，而这些元件中没有一个是用于Sm蛋白结合的。相反，包含Sm结合位点的U4小核仁RNA的3'末端结构域对于U4-U6相互作用和剪接体组装都是可有可无的。我们的结果支持将U4 snRNP组织成多个功能结构域，每个结构域在snRNP和剪接体组装的不同阶段发挥作用。
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