Novoyatleva Tatyana, Heinrich Bettina, Tang Yesheng, Benderska Natalya, Butchbach Matthew E R, Lorson Christian L, Lorson Monique A, Ben-Dov Claudia, Fehlbaum Pascale, Bracco Laurent, Burghes Arthur H M, Bollen Mathieu, Stamm Stefan
University of Erlangen, Institute for Biochemistry, Germany.
Hum Mol Genet. 2008 Jan 1;17(1):52-70. doi: 10.1093/hmg/ddm284. Epub 2007 Oct 2.
Alternative splicing emerges as one of the most important mechanisms to generate transcript diversity. It is regulated by the formation of protein complexes on pre-mRNA. We demonstrate that protein phosphatase 1 (PP1) binds to the splicing factor transformer2-beta1 (tra2-beta1) via a phylogenetically conserved RVDF sequence located on the RNA recognition motif (RRM) of tra2-beta1. PP1 binds directly to tra2-beta1 and dephosphorylates it, which regulates the interaction between tra2-beta1 and other proteins. Eight other proteins, including SF2/ASF and SRp30c, contain an evolutionary conserved PP1 docking motif in the beta-4 strand of their RRMs indicating that binding to PP1 is a new function of some RRMs. Reducing PP1 activity promotes usage of numerous alternative exons, demonstrating a role of PP1 activity in splice site selection. PP1 inhibition promotes inclusion of the survival of motoneuron 2 exon 7 in a mouse model expressing the human gene. This suggests that reducing PP1 activity could be a new therapeutic principle to treat spinal muscular atrophy and other diseases caused by missplicing events. Our data indicate that the binding of PP1 to evolutionary conserved motifs in several RRMs is the link between known signal transduction pathways regulating PP1 activity and pre-mRNA processing.
可变剪接成为产生转录本多样性的最重要机制之一。它由前体信使核糖核酸(pre-mRNA)上蛋白质复合物的形成所调控。我们证明蛋白磷酸酶1(PP1)通过位于tra2-β1的RNA识别基序(RRM)上的一个系统发育保守的RVDF序列与剪接因子transformer2-β1(tra2-β1)结合。PP1直接与tra2-β1结合并使其去磷酸化,从而调节tra2-β1与其他蛋白质之间的相互作用。包括SF2/ASF和SRp30c在内的其他8种蛋白质,在其RRM的β-4链中含有一个进化保守的PP1对接基序,这表明与PP1结合是一些RRM的新功能。降低PP1活性会促进众多可变外显子的使用,这表明PP1活性在剪接位点选择中发挥作用。在表达人类基因的小鼠模型中,PP1抑制促进运动神经元存活蛋白2外显子7的包含。这表明降低PP1活性可能是治疗脊髓性肌萎缩症和其他由错误剪接事件引起的疾病的新治疗原则。我们的数据表明,PP1与几个RRM中进化保守基序的结合是调节PP1活性的已知信号转导途径与前体信使核糖核酸加工之间的联系。