Hernández Félix, Pérez Mar, Lucas José J, Mata Ana M, Bhat Ratan, Avila Jesús
Centro de Biología Molecular Severo Ochoa Consejo Superior de Investigaciones Científicas/CSIC/Universidad Autónoma, Fac. Ciencias. Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain.
J Biol Chem. 2004 Jan 30;279(5):3801-6. doi: 10.1074/jbc.M311512200. Epub 2003 Nov 5.
Tauopathies, including Alzheimer's disease, are neurodegenerative disorders in which tau protein accumulates as a consequence of alterations in its metabolism. At least three different types of alterations have been described; in some cases, an aberrant mRNA splicing of tau exon 10 occurs; in other cases, the disorder is a consequence of missense mutations and, in most cases, aberrant tau hyperphosphorylation takes place. Glycogen synthase kinase-3 (GSK-3) has emerged as a key kinase that is able to interact with several proteins involved in the etiology of Alzheimer's disease and other tauopathies. Here, we have evaluated whether GSK-3 is also able to modulate tau-mRNA splicing. Our data demonstrate that GSK-3 inhibition in cultured neurons affects tau splicing resulting in an increase in tau mRNA containing exon 10. Pre-mRNA splicing is catalyzed by a multimolecular complex including members of the serine/arginine-rich (SR) family of splicing factors. Immunofluorescence studies showed that after GSK-3 inhibition, SC35, a member of the SR family, is redistributed and enriched in nuclear speckles and colocalizes with the kinase. Furthermore, immunoprecipitated SC35 is phosphorylated by recombinant GSK-3beta. Phosphorylation of a peptide from the SR domain by GSK-3 revealed that the peptide needs to be prephosphorylated, suggesting the involvement of a priming kinase. Our results demonstrate that GSK-3 plays a crucial role in tau exon 10 splicing, raising the possibility that GSK3 could contribute to tauopathies via aberrant tau splicing.
包括阿尔茨海默病在内的tau蛋白病是神经退行性疾病,其中tau蛋白由于其代谢改变而积累。至少已描述了三种不同类型的改变;在某些情况下,tau外显子10发生异常mRNA剪接;在其他情况下,该疾病是错义突变的结果,而在大多数情况下,发生异常的tau过度磷酸化。糖原合酶激酶-3(GSK-3)已成为一种关键激酶,能够与参与阿尔茨海默病和其他tau蛋白病病因的多种蛋白质相互作用。在此,我们评估了GSK-3是否也能够调节tau-mRNA剪接。我们的数据表明,在培养的神经元中抑制GSK-3会影响tau剪接,导致包含外显子10的tau mRNA增加。前体mRNA剪接由一个多分子复合物催化,该复合物包括富含丝氨酸/精氨酸(SR)的剪接因子家族成员。免疫荧光研究表明,在抑制GSK-3后,SR家族成员SC35重新分布并富集于核斑点中,并与该激酶共定位。此外,免疫沉淀的SC35被重组GSK-3β磷酸化。GSK-3对来自SR结构域的肽进行磷酸化表明该肽需要预先磷酸化,这表明存在引发激酶。我们的结果表明,GSK-3在tau外显子10剪接中起关键作用,这增加了GSK3可能通过异常的tau剪接导致tau蛋白病的可能性。