SOGA1和SOGA2/MTCL1是人类细胞中忠实的染色体分离所需的clasp相互作用蛋白。

IF 2.4 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Chromosome Research Pub Date : 2021-06-01 Epub Date: 2021-02-15 DOI:10.1007/s10577-021-09651-8
Luísa T Ferreira, Elsa Logarinho, Joana C Macedo, Ana Rita R Maia, Helder Maiato
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引用次数: 0

摘要

CLASPs是整个细胞周期中微管动力学的关键调节剂。在有丝分裂过程中,CLASPs独立地与生长的微管正端和着丝点相关联,并在染色体分离中发挥重要作用。在有丝分裂过程中对人类clasp1相互作用蛋白的蛋白质组学调查中,我们之前已经鉴定出SOGA1和SOGA2/MTCL1,其有丝分裂作用仍未被表征。在这里,我们进行了人类SOGA1和SOGA2/MTCL1在有丝分裂过程中的初步功能表征。利用针对SOGA蛋白的特异性多克隆抗体,我们证实了SOGA蛋白在有丝分裂过程中与CLASP1和CLASP2的表达和相互作用。此外,我们发现SOGA1和SOGA2/MTCL1在有丝分裂过程中都受到CDK1的磷酸化调控。免疫荧光分析显示,在整个有丝分裂过程中,SOGA2/MTCL1与纺锤体微管和纺锤杆共定位,并且在有丝分裂退出/细胞分裂期间,这两种SOGA蛋白都在中间体富集。gfp标记进一步揭示了SOGA2/MTCL1在着丝点上的微管独立定位。sirna介导的SOGA1和SOGA2/MTCL1敲低后的活细胞成像显示,它们在染色体分离的不同方面是独立必需的。因此,SOGA1和SOGA2/MTCL1是人类细胞有丝分裂过程中真正与clasp相互作用的蛋白,这是染色体分离所必需的。
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SOGA1 and SOGA2/MTCL1 are CLASP-interacting proteins required for faithful chromosome segregation in human cells.

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.

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来源期刊
Chromosome Research
Chromosome Research 生物-生化与分子生物学
CiteScore
4.70
自引率
3.80%
发文量
31
审稿时长
1 months
期刊介绍: Chromosome Research publishes manuscripts from work based on all organisms and encourages submissions in the following areas including, but not limited, to: · Chromosomes and their linkage to diseases; · Chromosome organization within the nucleus; · Chromatin biology (transcription, non-coding RNA, etc); · Chromosome structure, function and mechanics; · Chromosome and DNA repair; · Epigenetic chromosomal functions (centromeres, telomeres, replication, imprinting, dosage compensation, sex determination, chromosome remodeling); · Architectural/epigenomic organization of the genome; · Functional annotation of the genome; · Functional and comparative genomics in plants and animals; · Karyology studies that help resolve difficult taxonomic problems or that provide clues to fundamental mechanisms of genome and karyotype evolution in plants and animals; · Mitosis and Meiosis; · Cancer cytogenomics.
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