Kandala Divya T, Mohan Nimmy, A Vivekanand, A P Sudheesh, G Reshmi, Laishram Rakesh S
Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Trivandrum 695014, India.
Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Trivandrum 695014, India
Nucleic Acids Res. 2016 Jan 29;44(2):811-23. doi: 10.1093/nar/gkv1074. Epub 2015 Oct 22.
Almost all eukaryotic mRNAs have a poly (A) tail at the 3'-end. Canonical PAPs (PAPα/γ) polyadenylate nuclear pre-mRNAs. The recent identification of the non-canonical Star-PAP revealed specificity of nuclear PAPs for pre-mRNAs, yet the mechanism how Star-PAP selects mRNA targets is still elusive. Moreover, how Star-PAP target mRNAs having canonical AAUAAA signal are not regulated by PAPα is unclear. We investigate specificity mechanisms of Star-PAP that selects pre-mRNA targets for polyadenylation. Star-PAP assembles distinct 3'-end processing complex and controls pre-mRNAs independent of PAPα. We identified a Star-PAP recognition nucleotide motif and showed that suboptimal DSE on Star-PAP target pre-mRNA 3'-UTRs inhibit CstF-64 binding, thus preventing PAPα recruitment onto it. Altering 3'-UTR cis-elements on a Star-PAP target pre-mRNA can switch the regulatory PAP from Star-PAP to PAPα. Our results suggest a mechanism of poly (A) site selection that has potential implication on the regulation of alternative polyadenylation.
几乎所有真核生物的信使核糖核酸(mRNA)在3'端都有一个聚腺苷酸(poly (A))尾巴。典型的聚腺苷酸聚合酶(PAPα/γ)对细胞核内的前体mRNA进行聚腺苷酸化。最近发现的非典型的Star-PAP揭示了细胞核内聚腺苷酸聚合酶对前体mRNA的特异性,但Star-PAP选择mRNA靶标的机制仍不清楚。此外,尚不清楚Star-PAP如何靶向具有典型AAUAAA信号的mRNA且不受PAPα的调控。我们研究了Star-PAP选择前体mRNA靶标进行聚腺苷酸化的特异性机制。Star-PAP组装不同的3'端加工复合体,并独立于PAPα控制前体mRNA。我们鉴定出一个Star-PAP识别核苷酸基序,并表明Star-PAP靶标前体mRNA 3'非翻译区(UTR)上次优的下游序列元件(DSE)会抑制切割刺激因子64(CstF-64)的结合,从而阻止PAPα募集到其上。改变Star-PAP靶标前体mRNA上的3'-UTR顺式元件可以将调控性聚腺苷酸聚合酶从Star-PAP切换为PAPα。我们的结果提示了一种聚(A)位点选择机制,这对可变聚腺苷酸化的调控可能具有潜在意义。