Department of Biochemistry and Molecular Biology University of Miami Miller School of Medicine Miami FL, USA; Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, D.C, USA.
Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, D.C, USA.
Biochem Biophys Res Commun. 2024 Sep 3;723:150189. doi: 10.1016/j.bbrc.2024.150189. Epub 2024 May 29.
Casein kinase 1α (CK1α) is a serine/threonine protein kinase that acts in various cellular processes affecting cell division and signal transduction. CK1α is present as multiple splice variants that are distinguished by the presence or absence of a long insert (L-insert) and a short carboxyl-terminal insert (S-insert). When overexpressed, zebrafish CK1α splice variants exhibit different biological properties, such as subcellular localization and catalytic activity. However, whether endogenous, alternatively spliced CK1α gene products also differ in their biological functions has yet to be elucidated. Here, we identify a panel of splice variant specific CK1α antibodies and use them to show that four CK1α splice variants are expressed in mammals. We subsequently show that the relative abundance of CK1α splice variants varies across distinct mouse tissues and between various cancer cell lines. Furthermore, we identify pathways whose expression is noticeably altered in cell lines enriched with select splice variants of CK1α. Finally, we show that the S-insert of CK1α promotes the growth of HCT 116 cells as cells engineered to lack the S-insert display decreased cell growth. Together, we provide tools and methods to identify individual CK1α splice variants, which we use to begin to uncover the differential biological properties driven by specific splice variants of mammalian CK1α.
酪蛋白激酶 1α(CK1α)是一种丝氨酸/苏氨酸蛋白激酶,在影响细胞分裂和信号转导的各种细胞过程中发挥作用。CK1α 存在多种剪接变体,这些变体的区别在于是否存在长插入(L-插入)和短羧基末端插入(S-插入)。当过度表达时,斑马鱼 CK1α 剪接变体表现出不同的生物学特性,如亚细胞定位和催化活性。然而,内源性、选择性剪接的 CK1α 基因产物在其生物学功能上是否也存在差异,尚未阐明。在这里,我们鉴定了一组特异性 CK1α 剪接变体抗体,并利用它们表明哺乳动物中表达了四种 CK1α 剪接变体。随后,我们表明 CK1α 剪接变体的相对丰度在不同的小鼠组织和各种癌细胞系之间存在差异。此外,我们确定了表达明显改变的途径在富含 CK1α 选择性剪接变体的细胞系中。最后,我们表明 CK1α 的 S-插入促进 HCT 116 细胞的生长,因为缺乏 S-插入的细胞显示出细胞生长减少。总之,我们提供了识别单个 CK1α 剪接变体的工具和方法,我们利用这些工具和方法开始揭示由哺乳动物 CK1α 的特定剪接变体驱动的差异生物学特性。