Sierakowska H, Szer W, Furdon P J, Kole R
Nucleic Acids Res. 1986 Jul 11;14(13):5241-54. doi: 10.1093/nar/14.13.5241.
In vitro splicing of human beta-globin pre-mRNA can be fully inhibited by treatment of the splicing extract with polyclonal antibodies against hnRNP core proteins prior to the addition of pre-mRNA. Inhibition of the first step in the splicing pathway, cleavage at the 5' splice site and lariat formation, requires more antibodies than inhibition of the second step, cleavage at the 3' splice site and exon ligation. The anti-hnRNP antibodies can also inhibit the splicing reaction after the formation of the active nucleoprotein splicing complex which is known to occur during the initial lag period. Thus, hnRNP core proteins appear to be present in the complex that performs pre-mRNA splicing.
在添加前体mRNA之前,用人β-珠蛋白前体mRNA进行体外剪接时,通过用针对hnRNP核心蛋白的多克隆抗体处理剪接提取物,可完全抑制该剪接过程。与抑制第二步(3'剪接位点切割和外显子连接)相比,抑制剪接途径的第一步(5'剪接位点切割和套索形成)需要更多的抗体。抗hnRNP抗体也可以在活性核蛋白剪接复合物形成后抑制剪接反应,已知该复合物在初始延迟期形成。因此,hnRNP核心蛋白似乎存在于进行前体mRNA剪接的复合物中。