Department of Biological Sciences, University of Toledo, Toledo, OH, United States of America.
PLoS One. 2022 Jan 28;17(1):e0261098. doi: 10.1371/journal.pone.0261098. eCollection 2022.
JAK2 is cytokine-activated non-receptor tyrosine kinase. Although JAK2 is mainly localized at the plasma membrane, it is also present on the centrosome. In this study, we demonstrated that JAK2 localization to the centrosome depends on the SH2 domain and intact kinase activity. We created JAK2 mutants deficient in centrosomal localization ΔSH2, K882E and (ΔSH2, K882E). We showed that JAK2 WT clone strongly enhances cell proliferation as compared to control cells while JAK2 clones ΔSH2, K882E and (ΔSH2, K882E) proliferate slower than JAK2 WT cells. These mutant clones also progress much slower through the cell cycle as compared to JAK2 WT clone and the enhanced proliferation of JAK2 WT cells is accompanied by increased S -> G2 progression. Both the SH2 domain and the kinase activity of JAK2 play a role in prolactin-dependent activation of JAK2 substrate STAT5. We showed that JAK2 is an important regulator of centrosome function as the SH2 domain of JAK2 regulates centrosome amplification. The cells overexpressing ΔSH2 and (ΔSH2, K-E) JAK2 have almost three-fold the amplified centrosomes of WT cells. In contrast, the kinase activity of JAK2 is dispensable for centrosome amplification. Our observations provide novel insight into the role of SH2 domain and kinase activity of JAK2 in centrosome localization of JAK2 and in the regulation of cell growth and centrosome biogenesis.
JAK2 是细胞因子激活的非受体酪氨酸激酶。尽管 JAK2 主要定位于质膜,但它也存在于中心体上。在这项研究中,我们证明了 JAK2 向中心体的定位依赖于 SH2 结构域和完整的激酶活性。我们创建了缺乏中心体定位的 JAK2 突变体 ΔSH2、K882E 和(ΔSH2、K882E)。我们表明,与对照细胞相比,JAK2 WT 克隆强烈增强细胞增殖,而 JAK2 突变体 ΔSH2、K882E 和(ΔSH2、K882E)的增殖速度比 JAK2 WT 细胞慢。与 JAK2 WT 克隆相比,这些突变体克隆也更慢地通过细胞周期,并且 JAK2 WT 细胞的增强增殖伴随着 S->G2 进展的增加。JAK2 的 SH2 结构域和激酶活性都在催乳素依赖性 JAK2 底物 STAT5 的激活中起作用。我们表明 JAK2 是中心体功能的重要调节剂,因为 JAK2 的 SH2 结构域调节中心体扩增。过表达 ΔSH2 和(ΔSH2、K-E)JAK2 的细胞的扩增中心体几乎是 WT 细胞的三倍。相比之下,JAK2 的激酶活性对于中心体扩增是可有可无的。我们的观察结果为 JAK2 的 SH2 结构域和激酶活性在 JAK2 的中心体定位以及细胞生长和中心体发生的调节中的作用提供了新的见解。