Combined Inactivation of Pocket Proteins and APC/C<sup>Cdh1</sup> by Cdk4/6 Controls Recovery from DNA Damage in G1 Phase
Most Cyclin-dependent kinases (Cdks) are redundant for normal cell division. Here we tested whether these redundancies are maintained during cell cycle recovery after a DNA damage-induced arrest in G1. Using non-transformed RPE-1 cells, we find that while Cdk4 and Cdk6 act redundantly during normal...
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| Format: | Article |
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MDPI AG
2021-03-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/10/3/550 |
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| author | Indra A. Shaltiel Alba Llopis Melinda Aprelia Rob Klompmaker Apostolos Menegakis Lenno Krenning René H. Medema |
| author_facet | Indra A. Shaltiel Alba Llopis Melinda Aprelia Rob Klompmaker Apostolos Menegakis Lenno Krenning René H. Medema |
| author_sort | Indra A. Shaltiel |
| collection | DOAJ |
| description | Most Cyclin-dependent kinases (Cdks) are redundant for normal cell division. Here we tested whether these redundancies are maintained during cell cycle recovery after a DNA damage-induced arrest in G1. Using non-transformed RPE-1 cells, we find that while Cdk4 and Cdk6 act redundantly during normal S-phase entry, they both become essential for S-phase entry after DNA damage in G1. We show that this is due to a greater overall dependency for Cdk4/6 activity, rather than to independent functions of either kinase. In addition, we show that inactivation of pocket proteins is sufficient to overcome the inhibitory effects of complete Cdk4/6 inhibition in otherwise unperturbed cells, but that this cannot revert the effects of Cdk4/6 inhibition in DNA damaged cultures. Indeed, we could confirm that, in addition to inactivation of pocket proteins, Cdh1-dependent anaphase-promoting complex/cyclosome (APC/C<sup>Cdh1</sup>) activity needs to be inhibited to promote S-phase entry in damaged cultures. Collectively, our data indicate that DNA damage in G1 creates a unique situation where high levels of Cdk4/6 activity are required to inactivate pocket proteins and APC/C<sup>Cdh1</sup> to promote the transition from G1 to S phase. |
| first_indexed | 2024-03-09T05:36:41Z |
| format | Article |
| id | doaj.art-6239935eb2eb4a59bf037fae1993f10b |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T05:36:41Z |
| publishDate | 2021-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-6239935eb2eb4a59bf037fae1993f10b2023-12-03T12:28:33ZengMDPI AGCells2073-44092021-03-0110355010.3390/cells10030550Combined Inactivation of Pocket Proteins and APC/C<sup>Cdh1</sup> by Cdk4/6 Controls Recovery from DNA Damage in G1 PhaseIndra A. Shaltiel0Alba Llopis1Melinda Aprelia2Rob Klompmaker3Apostolos Menegakis4Lenno Krenning5René H. Medema6Division of Cell Biology and Cancer Genomics Center, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The NetherlandsDivision of Cell Biology and Cancer Genomics Center, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The NetherlandsDivision of Cell Biology and Cancer Genomics Center, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The NetherlandsDivision of Cell Biology and Cancer Genomics Center, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The NetherlandsDivision of Cell Biology and Cancer Genomics Center, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The NetherlandsDivision of Cell Biology and Cancer Genomics Center, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The NetherlandsDivision of Cell Biology and Cancer Genomics Center, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The NetherlandsMost Cyclin-dependent kinases (Cdks) are redundant for normal cell division. Here we tested whether these redundancies are maintained during cell cycle recovery after a DNA damage-induced arrest in G1. Using non-transformed RPE-1 cells, we find that while Cdk4 and Cdk6 act redundantly during normal S-phase entry, they both become essential for S-phase entry after DNA damage in G1. We show that this is due to a greater overall dependency for Cdk4/6 activity, rather than to independent functions of either kinase. In addition, we show that inactivation of pocket proteins is sufficient to overcome the inhibitory effects of complete Cdk4/6 inhibition in otherwise unperturbed cells, but that this cannot revert the effects of Cdk4/6 inhibition in DNA damaged cultures. Indeed, we could confirm that, in addition to inactivation of pocket proteins, Cdh1-dependent anaphase-promoting complex/cyclosome (APC/C<sup>Cdh1</sup>) activity needs to be inhibited to promote S-phase entry in damaged cultures. Collectively, our data indicate that DNA damage in G1 creates a unique situation where high levels of Cdk4/6 activity are required to inactivate pocket proteins and APC/C<sup>Cdh1</sup> to promote the transition from G1 to S phase.https://www.mdpi.com/2073-4409/10/3/550cell cyclecheckpointCDKsDNA damagerecoveryG1 |
| spellingShingle | Indra A. Shaltiel Alba Llopis Melinda Aprelia Rob Klompmaker Apostolos Menegakis Lenno Krenning René H. Medema Combined Inactivation of Pocket Proteins and APC/C<sup>Cdh1</sup> by Cdk4/6 Controls Recovery from DNA Damage in G1 Phase Cells cell cycle checkpoint CDKs DNA damage recovery G1 |
| title | Combined Inactivation of Pocket Proteins and APC/C<sup>Cdh1</sup> by Cdk4/6 Controls Recovery from DNA Damage in G1 Phase |
| title_full | Combined Inactivation of Pocket Proteins and APC/C<sup>Cdh1</sup> by Cdk4/6 Controls Recovery from DNA Damage in G1 Phase |
| title_fullStr | Combined Inactivation of Pocket Proteins and APC/C<sup>Cdh1</sup> by Cdk4/6 Controls Recovery from DNA Damage in G1 Phase |
| title_full_unstemmed | Combined Inactivation of Pocket Proteins and APC/C<sup>Cdh1</sup> by Cdk4/6 Controls Recovery from DNA Damage in G1 Phase |
| title_short | Combined Inactivation of Pocket Proteins and APC/C<sup>Cdh1</sup> by Cdk4/6 Controls Recovery from DNA Damage in G1 Phase |
| title_sort | combined inactivation of pocket proteins and apc c sup cdh1 sup by cdk4 6 controls recovery from dna damage in g1 phase |
| topic | cell cycle checkpoint CDKs DNA damage recovery G1 |
| url | https://www.mdpi.com/2073-4409/10/3/550 |
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