How Alterations in the Cdt1 Expression Lead to Gene Amplification in Breast Cancer
Abstract
We examined the effects of Cdt1 for gene amplification by using inducible system. We Obtained and validated reagents for inducible Cdt1-expression-system. Furthermore, we identified appropriated dose of MTX for testing gene amplification Cdt2 is a key molecule for inhibiting Cdt1 activity. The depletion of Cdt2 induces DNA re-replication in several cell lines. During experiment, we Identified a reduction of PCNA monoubiquitination in Cdt2 depleted MCF7 cells.Exogenous Cdt2 rescued the effect of si-Cdt2. Mutant of Cdt2 indicated that binding Cdt2 to Cul4/DDB1 was required for PCNA monoubiquitination. Furthermore, we identified that CRL4Cdt2 complex promotes PCNA-dependent translesion DNA synthesis.Additionally, we have identified p12 as a novel target of CRL4 complex. Cdt2 depletion canceled p12 degradation which regulated checkpoint pathway. Stable form of p12 promotes fork progression during intra-S-phase checkpoint.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 2011
- Accession Number
- ADA549648
Entities
People
- Kenta Terai
Organizations
- University of Virginia