Cdk1 phosphorylation negatively regulates the activity of Net1 towards RhoA during mitosis.
Cdk1 phosphorylation negatively regulates the activity of Net1 towards RhoA during mitosis.
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DOI：
10.1016/j.cellsig.2021.109926
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发表时间：
2021-04
影响因子：
4.8
通讯作者：
Frost JA
中科院分区：
文献类型：
--
作者：
Ulu A;Oh W;Zuo Y;Frost JA
神经上皮转化基因1（Net 1）是RhoA亚家族鸟嘌呤核苷酸交换因子，其在许多癌症中过表达并有助于癌细胞运动和增殖。Net 1在有丝分裂进程中也起着Rho GT3独立的作用，它促进Aurora A和Pak 2的中心体激活，并有助于前中期的染色体排列。为了了解控制Net 1有丝分裂功能的调控机制，我们研究了它是否被有丝分裂激酶Cdk 1磷酸化。我们观察到，Cdk 1磷酸化Net 1在其N-末端调控结构域和C-末端的多个位点在体外。通过对其中两个位点产生磷酸化特异性抗体，我们还证明了内源性和转染的Net 1在细胞中被Cdk 1磷酸化。用脂肪族或酸性残基取代主要Cdk 1磷酸化位点抑制Net 1与RhoA的相互作用，用Cdk 1抑制剂处理中期细胞增加Net 1活性。Cdk 1抑制也增加了Net 1定位到质膜和刺激皮质F-肌动蛋白的积累。此外，Net 1过表达导致纺锤体极性缺陷，其通过主要Cdk 1磷酸化位点的酸性取代而降低频率。这些数据表明，Cdk 1磷酸化Net 1在有丝分裂过程中，并建议，这负调控其信号的能力，以RhoA和改变肌动蛋白细胞骨架组织。
The Neuroepithelial transforming gene 1 (Net1) is a RhoA subfamily guanine nucleotide exchange factor that is overexpressed in a number of cancers and contributes to cancer cell motility and proliferation. Net1 also plays a Rho GTPase independent role in mitotic progression, where it promotes centrosomal activation of Aurora A and Pak2, and aids in chromosome alignment during prometaphase. To understand regulatory mechanisms controlling the mitotic function of Net1, we examined whether it was phosphorylated by the mitotic kinase Cdk1. We observed that Cdk1 phosphorylated Net1 on multiple sites in its N-terminal regulatory domain and C-terminus in vitro. By raising phospho-specific antibodies to two of these sites, we also demonstrated that both endogenous and transfected Net1 were phosphorylated by Cdk1 in cells. Substitution of the major Cdk1 phosphorylation sites with aliphatic or acidic residues inhibited the interaction of Net1 with RhoA, and treatment of metaphase cells with a Cdk1 inhibitor increased Net1 activity. Cdk1 inhibition also increased Net1 localization to the plasma membrane and stimulated cortical F-actin accumulation. Moreover, Net1 overexpression caused spindle polarity defects that were reduced in frequency by acidic substitution of the major Cdk1 phosphorylation sites. These data indicate that Cdk1 phosphorylates Net1 during mitosis and suggest that this negatively regulates its ability to signal to RhoA and alter actin cytoskeletal organization.
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影响因子：
8.8
作者：
通讯作者：
--
DOI：
10.1083/jcb.200207130
发表时间：
2003-01-20
期刊：
The Journal of cell biology
影响因子：
--
作者：
Maddox AS;Burridge K
通讯作者：
Burridge K
影响因子：
2.3
作者：
Enserink JM;Kolodner RD
通讯作者：
Kolodner RD
影响因子：
3.3
作者：
Menon, Sarita;Oh, Wonkyung;Frost, Jeffrey A.
通讯作者：
Frost, Jeffrey A.
影响因子：
5.3
作者：
Carr, Heather S.;Zuo, Yan;Frost, Jeffrey A.
通讯作者：
Frost, Jeffrey A.