CTCF is essential for proper mitotic spindle structure and anaphase segregation.
CTCF
DNA/chromatin
Mitosis
Mitotic spindle
Nucleus
Journal
Chromosoma
ISSN: 1432-0886
Titre abrégé: Chromosoma
Pays: Austria
ID NLM: 2985138R
Informations de publication
Date de publication:
20 Sep 2023
20 Sep 2023
Historique:
received:
11
01
2023
accepted:
08
09
2023
revised:
11
08
2023
medline:
20
9
2023
pubmed:
20
9
2023
entrez:
20
9
2023
Statut:
aheadofprint
Résumé
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.
Identifiants
pubmed: 37728741
doi: 10.1007/s00412-023-00810-w
pii: 10.1007/s00412-023-00810-w
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : NIGMS NIH HHS
ID : R00 GM123195
Pays : United States
Organisme : NHGRI NIH HHS
ID : UM1 HG011536
Pays : United States
Commentaires et corrections
Type : UpdateOf
Informations de copyright
© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.
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