Proliferating cell nuclear antigen interacts with the CRL4 ubiquitin ligase subunit CDT2 in DNA synthesis–induced degradation of CDT1

Feng, Lu; Saxena, Lovely; Hoang, Nam V.; Zhang, Chunxiao; Lee, Logan; Li, Wenjing; Gong, Xiaoshan; Lu, Fei; Sun, Hong; Zhang, Hui

doi:10.1074/jbc.ra118.003049

Cited by 16 publications

(17 citation statements)

References 26 publications

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“…Once this bistable switch is engaged, the persistent activity of CRL4 Cdt2 at aRCs during S phase generates hysteresis through the persistent degradation of p21, preventing it from interfering with DNA replication and insulating S phase progression from the influence of DNA damage . In addition to p21, it is likely that other cell cycle regulators that are degraded by CRL4 Cdt2 (e.g., Cdt1 ) should also exhibit hysteresis at the G1/S transition.…”

Section: Integration Of the Rb‐e2f Switch With Other Bistable Switchesmentioning

confidence: 99%

Bistable switches as integrators and actuators during cell cycle progression

et al. 2019

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Progression through the cell cycle is driven by bistable switches—specialized molecular circuits that govern transitions from one cellular state to another. Although the mechanics of bistable switches are relatively well understood, it is less clear how cells integrate multiple sources of molecular information to engage these switches. Here, we describe how bistable switches act as hubs of information processing and examine how variability, competition, and inheritance of molecular signals determine the timing of the Rb‐E2F bistable switch that controls cell cycle entry. Bistable switches confer both robustness and plasticity to cell cycle progression, ensuring that cell cycle events are performed completely and in the correct order, while still allowing flexibility to cope with ongoing stress and changing environmental conditions.

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Section: Integration Of the Rb‐e2f Switch With Other Bistable Switchesmentioning

confidence: 99%

Bistable switches as integrators and actuators during cell cycle progression

et al. 2019

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“…CRL4 DDB2 mediated degradation of DDB2 and p21 is PCNA-dependent, and PIP box contributes to the interaction between DDB2 and PCNA [ 41 ]. In addition, CDT2 is also a PIP box containing protein, and can directly bind to PCNA through the PIP box [ 6 , 42 ]. Our previous work showed that PIP box in CDT2 is essential for CDT1 degradation [ 42 ].…”

Section: Resultsmentioning

confidence: 99%

“…In addition, CDT2 is also a PIP box containing protein, and can directly bind to PCNA through the PIP box [ 6 , 42 ]. Our previous work showed that PIP box in CDT2 is essential for CDT1 degradation [ 42 ]. We supposed that the ubiquitination-mediated degradation of CDT2 by CRL4 DDB2 ligase is in a PCNA-dependent manner.…”

Section: Resultsmentioning

confidence: 99%

DDB2 regulates DNA replication through PCNA-independent degradation of CDT2

Zhang

et al. 2021

Cell Biosci

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Background Targeting ubiquitin-dependent proteolysis is one of the strategies in cancer therapy. CRLCDT2 and CRLDDB2 are two key E3 ubiquitin ligases involved in DNA replication and DNA damage repair. But CDT2 and DDB2 are opposite prognostic factors in kinds of cancers, and the underlining mechanism needs to be elucidated. Methods Small interfering RNAs were used to determine the function of target genes. Co-immunoprecipitation (Co-IP) was performed to detect the interaction between DDB2 and CDT2. Immunofluorescence assays and fluorescence activating cell sorting (FACS) were used to measure the change of DNA content. In vivo ubiquitination assay was carried out to clarify the ubiquitination of CDT2 mediated by DDB2. Cell synchronization was performed to arrest cells at G1/S and S phase. The mechanism involved in DDB2-mediated CDT2 degradation was investigated by constructing plasmids with mutant variants and measured by Western blot. Immunohistochemistry was performed to determine the relationship between DDB2 and CDT2. Paired two-side Student’s t-test was used to measure the significance of the difference between control group and experimental group. Results Knockdown of DDB2 stabilized CDT2, while over-expression of DDB2 enhanced ubiquitination of CDT2, and subsequentially degradation of CDT2. Although both DDB2 and CDT2 contain PIP (PCNA-interacting protein) box, PIP box is dispensable for DDB2-mediated CDT2 degradation. Knockdown of PCNA had negligible effects on the stability of CDT2, but promoted accumulation of CDT1, p21 and SET8. Silencing of DDB2 arrested cell cycle in G1 phase, destabilized CDT1 and reduced the chromatin loading of MCMs, thereby blocked the formation of polyploidy induced by ablation of CDT2. In breast cancer and ovarian teratoma tissues, high level of DDB2 was along with lower level of CDT2. Conclusions We found that CRL4DDB2 is the novel E3 ubiquitin ligases of CDT2, and DDB2 regulates DNA replication through indirectly regulates CDT1 protein stability by degrading CDT2 and promotes the assembly of pre-replication complex. Our results broaden the horizon for understanding the opposite function of CDT2 and DDB2 in tumorigenesis, and may provide clues for drug discovery in cancer therapy.

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“…In addition, CDT2 is also a PIP box containing protein, and can directly bind to PCNA through the PIP box [6,43]. Our previous work showed that PIP box in CDT2 is essential for CDT1 degradation [43]. We supposed that the ubiquitination-mediated degradation of CDT2 by CRL4 DDB2 ligase is in a PCNA-dependent manner.…”

Section: Ddb2 Interacts With Cdt2 and Mediates Its Ubiquitinationmentioning

confidence: 94%

“…The ubiquitination-mediated degradation of CDT2 by CRL4 DDB2 is independent of PCNA CRL4 DDB2 mediated degradation of DDB2 and p21 is PCNA-dependent, and PIP box contributes to the interaction between DDB2 and PCNA [42]. In addition, CDT2 is also a PIP box containing protein, and can directly bind to PCNA through the PIP box [6,43]. Our previous work showed that PIP box in CDT2 is essential for CDT1 degradation [43].…”

Section: Ddb2 Interacts With Cdt2 and Mediates Its Ubiquitinationmentioning

confidence: 97%

DDB2 Regulates DNA Replication through PCNA-Independent Degradation of CDT2

Zhang

et al. 2020

Preprint

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BackgroundTargeting ubiquitin-dependent proteolysis is one of the strategies in cancer therapy. CRLCDT2 and CRLDDB2 are two key E3 ubiquitin ligases involved in DNA replication and DNA damage repair. But CDT2 and DDB2 are opposite prognostic factors in kinds of cancers, and the underlining mechanism needs to be elucidated. MethodsSmall interfering RNAs were used to determine the function of target genes. Co-immunoprecipitation (Co-IP) was performed to detect the interaction between DDB2 and CDT2. Immunofluorescence assays and fluorescence activating cell sorting (FACS) were used to measure the change of DNA content. In vivo ubiquitination assay was carried out to clarify the ubiquitination of CDT2 mediated by DDB2. Cell synchronization was performed to arrest cells at G1/S and S phase. The mechanism involved in DDB2-mediated CDT2 degradation was investigated by constructing plasmids with mutant variants and measured by Western blot. Immunohistochemistry was performed to determine the relationship between DDB2 and CDT2. Paired two-side Student’s t-test was used to measure the signiﬁcance of the difference between control group and experimental group.Results Knockdown of DDB2 stabilized CDT2, while over-expression of DDB2 enhanced ubiquitination of CDT2, and subsequentially degradation of CDT2. Although both DDB2 and CDT2 contain PIP (PCNA-interacting protein) box, PIP box is dispensable for DDB2-mediated CDT2 degradation. Knockdown of PCNA had negligible effects on the stability of CDT2, but promoted accumulation of CDT1, p21 and SET8. Silencing of DDB2 arrested cell cycle in G1 phase, destabilized CDT1 and reduced the chromatin loading of MCMs, thereby blocked the formation of polyploidy induced by ablation of CDT2. In breast cancer and ovarian teratoma tissues, high level of DDB2 was along with lower level of CDT2.ConclusionWe found that CRL4DDB2 is the novel E3 ubiquitin ligases of CDT2, and DDB2 regulates DNA replication through indirectly regulates CDT1 protein stability by degrading CDT2 and promotes the assembly of pre-replication complex. Our results broaden the horizon for understanding the opposite function of CDT2 and DDB2 in tumorigenesis, and may provide clues for drug discovery in cancer therapy.

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Proliferating cell nuclear antigen interacts with the CRL4 ubiquitin ligase subunit CDT2 in DNA synthesis–induced degradation of CDT1

Cited by 16 publications

References 26 publications

Bistable switches as integrators and actuators during cell cycle progression

Bistable switches as integrators and actuators during cell cycle progression

DDB2 regulates DNA replication through PCNA-independent degradation of CDT2

DDB2 Regulates DNA Replication through PCNA-Independent Degradation of CDT2

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