Niinuma Sho, Fukaya Takashi, Tomari Yukihide
Institute of Molecular and Cellular Biosciences, The University of Tokyo, Bunkyo-ku, Tokyo 113-0032, JapanDepartment of Computational Biology and Medical Sciences, The University of Tokyo, Bunkyo-ku, Tokyo 113-0032, Japan.
RNA. 2016 Oct;22(10):1550-9. doi: 10.1261/rna.057679.116. Epub 2016 Aug 2.
MicroRNAs (miRNAs) recruit the CCR4-NOT complex, which contains two deadenylases, CCR4 and CAF1, to promote shortening of the poly(A) tail. Although both CCR4 and CAF1 generally have a strong preference for poly(A) RNA substrates, it has been reported from yeast to humans that they can also remove non-A residues in vitro to various degrees. However, it remains unknown how CCR4 and CAF1 remove non-A sequences. Herein we show that Drosophila miRNAs can promote the removal of 3'-terminal non-A residues in an exonucleolytic manner, but only if an upstream poly(A) sequence exists. This non-A removing reaction is directly catalyzed by both CCR4 and CAF1 and depends on the balance between the length of the internal poly(A) sequence and that of the downstream non-A sequence. These results suggest that the CCR4-NOT complex has an intrinsic activity to remove the 3'-terminal non-A modifications downstream from the poly(A) tail.
微小RNA(miRNA)招募CCR4-NOT复合物,该复合物包含两种去腺苷酸化酶CCR4和CAF1,以促进多聚腺苷酸(poly(A))尾巴缩短。尽管CCR4和CAF1通常对poly(A) RNA底物有很强的偏好,但从酵母到人类都有报道称它们在体外也能不同程度地去除非A残基。然而,CCR4和CAF1如何去除非A序列仍不清楚。在此我们表明,果蝇miRNA可以以外切核酸酶的方式促进3'末端非A残基的去除,但前提是存在上游poly(A)序列。这种非A去除反应由CCR4和CAF1直接催化,并取决于内部poly(A)序列长度与下游非A序列长度之间的平衡。这些结果表明,CCR4-NOT复合物具有去除poly(A)尾巴下游3'末端非A修饰的内在活性。