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Excessive E2F Transcription in Single Cancer Cells Precludes Transient Cell Cycle Exit after DNA Damage

29 Pages Posted: 20 Apr 2020 Sneak Peek Status: Under Review

See all articles by Hendrika A. Segeren

Hendrika A. Segeren

Utrecht University - Department of Biomolecular Health Sciences

Lotte van Rijnberk

Utrecht University - Department of Biomolecular Health Sciences

Eva Moreno

Utrecht University - Department of Biomolecular Health Sciences

Frank Riemers

Utrecht University - Department of Biomolecular Health Sciences

Ruixue Yuan

Utrecht University - Department of Biomolecular Health Sciences

Richard Wubbolts

Utrecht University - Department of Biomolecular Health Sciences

Alain de Bruin

Utrecht University - Department of Biomolecular Health Sciences

Bart Westendorp

Utrecht University - Department of Biomolecular Health Sciences

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Abstract

E2F transcription factors control the expression of a large network of cell cycle genes and are essential for S-phase entry. Cancers often show upregulation of E2F target gene expression, which can be partially explained by an increased percentage of replicating cells. However, we now demonstrate in human cancer biopsies that individual cycling neoplastic cells display abnormally high levels of E2F-dependent transcription using single cell RNA sequencing. Since cancer patients are treated with DNA damaging drugs, we determined whether elevated E2F activity has impact on the response to DNA damage. We specifically increased the expression of E2F target genes during S and G2-phase by deleting the atypical E2F transcriptional repressors (E2F7/8) in untransformed cells. Live cell imaging revealed that cells with elevated E2F activity in S-G2 failed to arrest and underwent unscheduled mitosis after neocarzinostatin-induced DNA damage. In contrast, cells with physiological E2F-activity completed S-phase and subsequently exit the cell cycle in response to DNA damage. This exit into a G1-like arrest is dependent on transcriptional repression of the E2F-target Emi1 resulting in activation of APC/CCdh1. Interestingly, ~30% of these arrested 4N-G1 cells could eventually inactivate APC/CCdh1 to execute a second round of DNA replication and mitosis, resulting in the formation of tetraploid cells. In summary, we show that a subpopulation of cancer cells display extreme high E2F activity. This unrestricted E2F activity can prevent cell cycle exit after DNA damage and likely increases the chance to acquire more genetic changes and therefore potentially promote malignant progression and reduce sensitivity to anti-cancer drugs.

Keywords: Cell Cycle, Anaphase-Promoting Complex/Cyclosome, E2F transcription, Live cell imaging, Single Cell Sequencing, DNA damage, cancer, P53, Emi1, Tetraploidy

Suggested Citation

Segeren, Hendrika A. and van Rijnberk, Lotte and Moreno, Eva and Riemers, Frank and Yuan, Ruixue and Wubbolts, Richard and de Bruin, Alain and Westendorp, Bart, Excessive E2F Transcription in Single Cancer Cells Precludes Transient Cell Cycle Exit after DNA Damage. Available at SSRN: https://ssrn.com/abstract=3570558 or http://dx.doi.org/10.2139/ssrn.3570558
This is a paper under consideration at Cell Press and has not been peer-reviewed.

Hendrika A. Segeren

Utrecht University - Department of Biomolecular Health Sciences

Netherlands

Lotte Van Rijnberk

Utrecht University - Department of Biomolecular Health Sciences

Netherlands

Eva Moreno

Utrecht University - Department of Biomolecular Health Sciences

Netherlands

Frank Riemers

Utrecht University - Department of Biomolecular Health Sciences

Netherlands

Ruixue Yuan

Utrecht University - Department of Biomolecular Health Sciences

Netherlands

Richard Wubbolts

Utrecht University - Department of Biomolecular Health Sciences

Netherlands

Alain De Bruin

Utrecht University - Department of Biomolecular Health Sciences

Netherlands

Bart Westendorp (Contact Author)

Utrecht University - Department of Biomolecular Health Sciences ( email )

Netherlands

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