CTCF is Essential for Proper Mitotic Spindle Structure and Anaphase Segregation

Overview

Journal bioRxiv

Date 2023 Jan 30

PMID 36712070

Authors

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

Affiliations

Soon will be listed here.

Abstract

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 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. 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 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.

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