Topoisomerase IIA is essential for maintenance of mitotic chromosome structure

Data files for the manuscript #2020-01760 "Topoisomerase IIA is essential for maintenance of mitotic chromosome structure" in press in the journal "Preceedings of the National Academy of Sciences" (PNAS).

Abstract

Topoisomerase IIa (TOP2A) is a core component of mitotic chromosomes and important for establishing mitotic chromosome condensation. The primary roles of TOP2A in mitosis have been difficult to decipher due to its multiple functions across the cell cycle. To more precisely understand the role of TOP2A in mitosis, we used the auxin inducible degron (AID) system to rapidly degrade the protein at different stages of the human cell cycle. Removal of TOP2A prior to mitosis does not affect prophase timing or the initiation of chromosome condensation. Instead, it prevents chromatin condensation in prometaphase, extends the length of prometaphase and ultimately causes cells to exit mitosis without chromosome segregation occurring. Surprisingly, we find that removal of TOP2A from cells arrested in prometaphase or metaphase cause dramatic loss of compacted mitotic chromosome structure and conclude that TOP2A is crucial for maintenance of mitotic chromosomes. Treatments with drugs used to poison/inhibit TOP2A function, such as etoposide and ICRF-193, does not phenocopy the effects on chromosome structure of TOP2A degradation by AID. Our data, points to a role for TOP2A as a structural chromosome maintenance enzyme component locking in condensation states once sufficient compaction is achieved.