Laboratory of Biochemistry, Department of Chemistry, Aristotle University, 54124 Thessaloniki, Greece.
Department of Neurobiology, Care Sciences and Society, Division of Neurogeriatrics, Center for Alzheimer Research, Karolinska Institutet, 10435 Stockholm, Sweden.
Cells. 2022 Dec 28;12(1):126. doi: 10.3390/cells12010126.
Although SRPKs were discovered nearly 30 years ago, our understanding of their mode of regulation is still limited. Regarded as constitutively active enzymes known to participate in diverse biological processes, their prominent mode of regulation mainly depends on their intracellular localization. Molecular chaperones associate with a large internal spacer sequence that separates the bipartite kinase catalytic core and modulates the kinases' partitioning between the cytoplasm and nucleus. Besides molecular chaperones that function as anchoring proteins, a few other proteins were shown to interact directly with SRPK1, the most-studied member of SRPKs, and alter its activity. In this study, we identified TAF15, which has been involved in transcription initiation, splicing, DNA repair, and RNA maturation, as a novel SRPK1-interacting protein. The C-terminal RGG domain of TAF15 was able to associate with SRPK1 and downregulate its activity. Furthermore, overexpression of this domain partially relocalized SRPK1 to the nucleus and resulted in hypophosphorylation of SR proteins, inhibition of splicing of a reporter minigene, and inhibition of Lamin B receptor phosphorylation. We further demonstrated that peptides comprising the RGG repeats of nucleolin, HNRPU, and HNRNPA2B1, were also able to inhibit SRPK1 activity, suggesting that negative regulation of SRPK1 activity might be a key biochemical property of RGG motif-containing proteins.
虽然 SRPKs 是在近 30 年前发现的,但我们对其调节方式的理解仍然有限。作为已知参与多种生物过程的组成性活性酶,它们的主要调节方式主要取决于其细胞内定位。分子伴侣与一个大的内部间隔序列相关联,该间隔序列将二部分激酶催化核心分开,并调节激酶在细胞质和细胞核之间的分配。除了作为锚定蛋白发挥作用的分子伴侣外,还有一些其他蛋白质被证明可以直接与 SRPK1 相互作用,SRPK1 是 SRPKs 中研究最多的成员,并改变其活性。在这项研究中,我们鉴定了 TAF15,它已参与转录起始、剪接、DNA 修复和 RNA 成熟,是一种新型的 SRPK1 相互作用蛋白。TAF15 的 C 末端 RGG 结构域能够与 SRPK1 结合并下调其活性。此外,该结构域的过表达部分将 SRPK1 重新定位到细胞核,并导致 SR 蛋白的低磷酸化、报告基因 minigene 的剪接抑制以及 lamin B 受体磷酸化的抑制。我们进一步证明,核仁蛋白、HNRPU 和 HNRNPA2B1 的 RGG 重复肽也能够抑制 SRPK1 活性,这表明 RGG 基序富含蛋白质的 SRPK1 活性的负调节可能是一种关键的生化特性。