期刊
JOURNAL OF CELL SCIENCE
卷 131, 期 4, 页码 -出版社
COMPANY BIOLOGISTS LTD
DOI: 10.1242/jcs.210187
关键词
CDK1; Chromosome; DT40; Mitosis; Auxin inducible degron system; Condensin
类别
资金
- Wellcome Trust [107022, 203149]
- Japan Society for the Promotion of Science [16K15095]
- Mochida Memorial Foundation for Medical and Pharmaceutical Research, SGH Foundation, Japan
- Sumitomo Foundation
- Cannon Foundation
- Biotechnology and Biological Sciences Research Council [BB/L014416/1] Funding Source: researchfish
- BBSRC [BB/L014416/1] Funding Source: UKRI
- Grants-in-Aid for Scientific Research [16K15095] Funding Source: KAKEN
The requirement for condensin in chromosome formation in somatic cells remains unclear, as imperfectly condensed chromosomes do form in cells depleted of condensin by conventional methodologies. In order to dissect the roles of condensin at different stages of vertebrate mitosis, we have established a versatile cellular system that combines auxin-mediated rapid degradation with chemical genetics to obtain near-synchronous mitotic entry of chicken DT40 cells in the presence and absence of condensin. We analyzed the outcome by live-and fixed-cell microscopy methods, including serial block face scanning electron microscopy with digital reconstruction. Following rapid depletion of condensin, chromosomal defects were much more obvious than those seen after a slow depletion of condensin. The total mitotic chromatin volume was similar to that in control cells, but a single mass of mitotic chromosomes was clustered at one side of a bent mitotic spindle. Cultures arrest at prometaphase, eventually exiting mitosis without segregating chromosomes. Experiments where the auxin concentration was titrated showed that different condensin levels are required for anaphase chromosome segregation and formation of a normal chromosome architecture.
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