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Temperature-Induced Uncoupling of Cell Cycle Regulators

Hanieh Falahati, Woonyung Hur, Stefano Di Talia, View ORCID ProfileEric F. Wieschaus
doi: https://doi.org/10.1101/2020.02.11.943266
Hanieh Falahati
1Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ 08544
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Woonyung Hur
2Department of Cell Biology, Duke University Medical Center, Durham, NC 27710
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Stefano Di Talia
2Department of Cell Biology, Duke University Medical Center, Durham, NC 27710
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Eric F. Wieschaus
1Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ 08544
3Howard Hughes Medical Institute, Department of Molecular Biology, Princeton University, Princeton, NJ 08544
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  • ORCID record for Eric F. Wieschaus
  • For correspondence: efw@princeton.edu
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Abstract

While feedback loops are essential for robustness in signaling systems, they make discerning the role of individual components challenging. Here we introduce temperature as a powerful perturbation method for uncoupling enzymatic processes, by exposing the differential sensitivity of limiting reactions to temperature due to their activation energies. Using this method, we study the sensitivity to temperature of different cell cycle events of early fly embryos. While the subdivision of cell cycle steps is conserved across a wide range of temperatures (5-35°C), the transition into prometaphase exhibits the most sensitivity, arguing that it has a different mechanism of regulation. Using a biosensor, we quantify the activity of Cdk1 and show that the activation of Cdk1 drives entry into prometaphase but is not required for earlier events. In fact, chromosome condensation can still occur when Cdk1 is inhibited pharmacologically. These results demonstrate that different kinases are rate-limiting for different steps of mitosis.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted February 11, 2020.
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Temperature-Induced Uncoupling of Cell Cycle Regulators
Hanieh Falahati, Woonyung Hur, Stefano Di Talia, Eric F. Wieschaus
bioRxiv 2020.02.11.943266; doi: https://doi.org/10.1101/2020.02.11.943266
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Temperature-Induced Uncoupling of Cell Cycle Regulators
Hanieh Falahati, Woonyung Hur, Stefano Di Talia, Eric F. Wieschaus
bioRxiv 2020.02.11.943266; doi: https://doi.org/10.1101/2020.02.11.943266

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