CTCF对于正确的有丝分裂纺锤体结构和后期分离是必不可少的。

IF 2.5 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Chromosoma Pub Date : 2024-07-01 Epub Date: 2023-09-20 DOI:10.1007/s00412-023-00810-w
Katherine Chiu, Yasmin Berrada, Nebiyat Eskndir, Dasol Song, Claire Fong, Sarah Naughton, Tina Chen, Savanna Moy, Sarah Gyurmey, Liam James, Chimere Ezeiruaku, Caroline Capistran, Daniel Lowey, Vedang Diwanji, Samantha Peterson, Harshini Parakh, Ayanna R Burgess, Cassandra Probert, Annie Zhu, Bryn Anderson, Nehora Levi, Gabi Gerlitz, Mary C Packard, Katherine A Dorfman, Michael Seifu Bahiru, Andrew D Stephens
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引用次数: 0

摘要

有丝分裂是一个重要的过程,在这个过程中,复制的基因组被平等地分离成两个子细胞。CTCF已被报道存在于有丝分裂中,并在定位CENP-E中发挥作用,但其对有丝分裂保真度的重要性仍有待确定。为了评估CTCF在有丝分裂中的重要性,我们跟踪了野生型和两种不同的基于CTCF CRISPR的基因敲除中的有丝分裂行为。我们发现,通过SiR-DNA的延时成像,敲低CTCF会导致有丝分裂延长和后期分离失败。CTCF敲除不改变细胞周期或有丝分裂检查点,后者在诺可达唑治疗后被激活。CTCF敲除中有丝分裂纺锤体的免疫荧光成像显示,通过三极/四极纺锤体和纺锤体极后面的染色体出现紊乱。间期细胞核的成像显示细胞核大小急剧增加,这与后期未能分割重复的基因组一致。通过GSK923295对CNEP-E的长期抑制概括了CTCF敲低具有极性染色体的异常有丝分裂纺锤体和增加的细胞核大小。相对于野生型,CTCF敲除中细胞核形状的群体测量没有显示圆形度降低或细胞核起泡增加。然而,与成功的有丝分裂相比,失败的有丝裂确实显示出异常的核形态,这表明群体图像不能捕捉到个体的行为。因此,CTCF对适当的中期组织和后期分离都很重要,后期分离可能通过其在招募CENP-E中的已知作用影响相间核的大小和形状。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

CTCF is essential for proper mitotic spindle structure and anaphase segregation.

CTCF is essential for proper mitotic spindle structure and anaphase segregation.

Mitosis is an essential process in which the duplicated genome is segregated equally into two daughter cells. CTCF has been reported to be present in mitosis and has a role in localizing CENP-E, but its importance for mitotic fidelity remains to be determined. To evaluate the importance of CTCF in mitosis, we tracked mitotic behaviors in wild-type and two different CTCF CRISPR-based genetic knockdowns. We find that knockdown of CTCF results in prolonged mitoses and failed anaphase segregation via time-lapse imaging of SiR-DNA. CTCF knockdown did not alter cell cycling or the mitotic checkpoint, which was activated upon nocodazole treatment. Immunofluorescence imaging of the mitotic spindle in CTCF knockdowns revealed disorganization via tri/tetrapolar spindles and chromosomes behind the spindle pole. Imaging of interphase nuclei showed that nuclear size increased drastically, consistent with failure to divide the duplicated genome in anaphase. Long-term inhibition of CNEP-E via GSK923295 recapitulates CTCF knockdown abnormal mitotic spindles with polar chromosomes and increased nuclear sizes. Population measurements of nuclear shape in CTCF knockdowns do not display decreased circularity or increased nuclear blebbing relative to wild-type. However, failed mitoses do display abnormal nuclear morphologies relative to successful mitoses, suggesting that population images do not capture individual behaviors. Thus, CTCF is important for both proper metaphase organization and anaphase segregation which impacts the size and shape of the interphase nucleus likely through its known role in recruiting CENP-E.

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来源期刊
Chromosoma
Chromosoma 生物-生化与分子生物学
CiteScore
3.30
自引率
6.20%
发文量
17
审稿时长
1 months
期刊介绍: Chromosoma publishes research and review articles on the functional organization of the eukaryotic cell nucleus, with a particular emphasis on the structure and dynamics of chromatin and chromosomes; the expression and replication of genomes; genome organization and evolution; the segregation of genomes during meiosis and mitosis; the function and dynamics of subnuclear compartments; the nuclear envelope and nucleocytoplasmic interactions, and more. The scope of Chromosoma encompasses genetic, biophysical, molecular and cell biological studies. Average time from receipt of contributions to first decision: 22 days Publishes research and review articles on the functional organization of the eukaryotic cell nucleus Topics include structure and dynamics of chromatin and chromosomes; the expression and replication of genomes; genome organization and evolution; the segregation of genomes during meiosis and mitosis and more Encompasses genetic, biophysical, molecular and cell biological studies.
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