Thomé Vanessa, Ferreira Pedro B, Lubini Greice, Nogueira Fernanda M, Strini Edward J, Pinoti Vitor F, Cruz Joelma O, San Martin Juca A B, Quiapim Andréa C, daSilva Luis L P, Goldman Maria Helena S
Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto 14040-901, SP, Brazil.
PPG Genética, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto 14001-970, SP, Brazil.
Int J Mol Sci. 2024 Dec 24;26(1):46. doi: 10.3390/ijms26010046.
In the flower development study, we identified SCI1 (Stigma/style Cell-cycle Inhibitor 1), a regulator of cell proliferation. SCI1 interacts with NtCDKG;2 ( Cyclin-Dependent Kinase G;2), a homolog of human CDK11, which is responsible for RanGTP-dependent microtubule stabilization, regulating spindle assembly rate. In a Y2H screening of a cDNA library using NtCDKG;2 as bait, a RanBP1 (Ran-Binding Protein 1) was revealed as its interaction partner. RanBP1 is an essential regulatory protein of the RanGTPase system, contributing to the formation of the Ran gradient, which modulates different important cellular processes. RanBP1 is crucial in the nuclear import/export machinery during interphase and spindle checkpoint formation during cell division. These processes are well studied in animals, but very little is known about them in plants. We confirmed NtCDKG;2 and NtRanBP1 interaction by pairwise Y2H and characterized the localization of both proteins during plant cell division. We demonstrated the presence of NtRanBP1 in the cytoplasm during interphase and its nuclear arrest at mitosis onset. Meanwhile, we showed that NtCDKG;2 is localized in the mitotic spindle during cell division, indicating an analogous function to the human CDK11. We propose that the phosphorylation of the nuclear export signal at RanBP1 by NtCDKG;2 may be responsible for the reported nuclear arrest.
在花发育研究中,我们鉴定出了SCI1(柱头/花柱细胞周期抑制剂1),一种细胞增殖调节因子。SCI1与NtCDKG;2(细胞周期蛋白依赖性激酶G;2)相互作用,NtCDKG;2是人类CDK11的同源物,负责RanGTP依赖性微管稳定,调节纺锤体组装速率。在用NtCDKG;2作为诱饵对cDNA文库进行的酵母双杂交筛选中,发现RanBP1(Ran结合蛋白1)是其相互作用伙伴。RanBP1是RanGTPase系统的一种重要调节蛋白,有助于形成Ran梯度,从而调节不同的重要细胞过程。RanBP1在间期的核输入/输出机制以及细胞分裂期间的纺锤体检查点形成中至关重要。这些过程在动物中已得到充分研究,但在植物中却知之甚少。我们通过成对酵母双杂交证实了NtCDKG;2和NtRanBP1的相互作用,并对这两种蛋白在植物细胞分裂过程中的定位进行了表征。我们证明了NtRanBP1在间期存在于细胞质中,在有丝分裂开始时被阻滞在细胞核中。同时,我们表明NtCDKG;2在细胞分裂期间定位于有丝分裂纺锤体中,表明其与人类CDK11具有类似功能。我们提出,NtCDKG;2对RanBP1核输出信号的磷酸化可能是导致所报道的核阻滞的原因。