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SOGA1 和 SOGA2/MTCL1 是与 CLASP 相互作用的蛋白质,对于人类细胞中染色体的正确分离是必需的。

SOGA1 and SOGA2/MTCL1 are CLASP-interacting proteins required for faithful chromosome segregation in human cells.

机构信息

Chromosome Instability & Dynamics Group, i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen 208, 4200-135, Porto, Portugal.

Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua Alfredo Allen 208, 4200-135, Porto, Portugal.

出版信息

Chromosome Res. 2021 Jun;29(2):159-173. doi: 10.1007/s10577-021-09651-8. Epub 2021 Feb 15.

Abstract

CLASPs are key modulators of microtubule dynamics throughout the cell cycle. During mitosis, CLASPs independently associate with growing microtubule plus-ends and kinetochores and play essential roles in chromosome segregation. In a proteomic survey for human CLASP1-interacting proteins during mitosis, we have previously identified SOGA1 and SOGA2/MTCL1, whose mitotic roles remained uncharacterized. Here we performed an initial functional characterization of human SOGA1 and SOGA2/MTCL1 during mitosis. Using specific polyclonal antibodies raised against SOGA proteins, we confirmed their expression and reciprocal interaction with CLASP1 and CLASP2 during mitosis. In addition, we found that both SOGA1 and SOGA2/MTCL1 are phospho-regulated during mitosis by CDK1. Immunofluorescence analysis revealed that SOGA2/MTCL1 co-localizes with mitotic spindle microtubules and spindle poles throughout mitosis and both SOGA proteins are enriched at the midbody during mitotic exit/cytokinesis. GFP-tagging of SOGA2/MTCL1 further revealed a microtubule-independent localization at kinetochores. Live-cell imaging after siRNA-mediated knockdown of SOGA1 and SOGA2/MTCL1 showed that they are independently required for distinct aspects of chromosome segregation. Thus, SOGA1 and SOGA2/MTCL1 are bona fide CLASP-interacting proteins during mitosis required for faithful chromosome segregation in human cells.

摘要

CLASPs 是整个细胞周期中小管动力学的关键调节剂。在有丝分裂过程中,CLASPs 独立地与生长中的微管正极和动粒结合,并在染色体分离中发挥重要作用。在一项针对人类 CLASP1 有丝分裂相关蛋白的蛋白质组学调查中,我们之前已经鉴定了 SOGA1 和 SOGA2/MTCL1,但其有丝分裂作用仍未被阐明。在这里,我们对人类 SOGA1 和 SOGA2/MTCL1 在有丝分裂过程中的初始功能进行了表征。使用针对 SOGA 蛋白特异性的多克隆抗体,我们证实了它们在有丝分裂过程中的表达以及与 CLASP1 和 CLASP2 的相互作用。此外,我们发现 SOGA1 和 SOGA2/MTCL1 在有丝分裂过程中都受到 CDK1 的磷酸化调节。免疫荧光分析显示,SOGA2/MTCL1 在整个有丝分裂过程中与有丝分裂纺锤体微管和纺锤体极共定位,并且在有丝分裂退出/胞质分裂期间,这两种 SOGA 蛋白都在中体处富集。SOGA2/MTCL1 的 GFP 标记进一步显示了在动粒处的微管非依赖性定位。在 siRNA 介导的 SOGA1 和 SOGA2/MTCL1 敲低后进行活细胞成像显示,它们独立地参与了染色体分离的不同方面。因此,SOGA1 和 SOGA2/MTCL1 是有丝分裂过程中真正的 CLASP 相互作用蛋白,对于人类细胞中忠实的染色体分离是必需的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a2c/7610860/433a19edeed7/EMS116943-f001.jpg

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