- Poster presentation
Regulation of cyclin D1by the BRCA1–BARD1 complex
Breast Cancer Research volume 10, Article number: P22 (2008)
BRCA1 and cyclin D1 are both essential for normal breast development and mutation or aberration of their expression is associated with breast cancer [1, 2]. Cyclin D1 is best known as a G1 cyclin where it regulates the G1 to S phase transition by acting as a rate-limiting subunit of CDK4/6 kinase activity. More recently, however, Stacey has demonstrated that cyclin D1 levels in G2/M determine whether a cell continues to proliferate or exits the cell cycle . The majority of BRCA1 in the cell is bound to BARD1 through their N-terminal RING domains. Hetero-dimerization is essential for the stability and correct localization of the complex and confers ubiquitin ligase activity to BRCA1. The importance of the ligase activity of BRCA1 to breast cancer development is inferred from the fact that N-terminal disease-associated mutations are proposed to reduce ligase activity .
Protein–protein interactions were demonstrated using yeast-two-hybrid and coimmunoprecipitation. Protein levels were altered through overexpression, siRNA and antisense technology. The effect of proteasome inhibitors and cycloheximide treatment was also examined.
We initially identified cyclin D1 as a binding partner of BARD1 in a yeast-two-hybrid screen and defined the minimal binding region as the N-terminus of BARD1. This interaction was confirmed in vivo by coimmunoprecipitation. The N-terminus of BARD1 also binds BRCA1 and imparts ubiquitin ligase activity to the complex. Covalent modification of proteins with ubiquitin is a common regulatory mechanism in eukaryotic cells. Traditionally, polyubiquitin chains linked through lysine 48 target proteins for degradation by the 26 S proteasome. We have demonstrated that cyclin D1 protein levels are inversely related to BRCA1 and BARD1 levels in several model systems. Furthermore, regulation of cyclin D1 levels occurs through a post-transcriptional mechanism and requires the ligase activity of BRCA1. Interestingly, this phenomenon is cell-cycle regulated, occurring in G2/M.
We propose that cyclin D1 is a potential substrate for BRCA1 ubiquitination and that this targets cyclin D1 for proteasomal-mediated degradation. Future work will focus on ascertaining the functional consequence of cyclin D1 regulation by the BRCA1–BARD1 complex; in particular, the impact of BRCA1, mediated through regulation of cyclin D1, on the proliferation versus differentiation decision.
Sicinski P, Donaher JL, Parker SB, Li T, Fazeli A, Gardner H, Haslam SZ, Bronson RT, Elledge SJ, Weinberg RA: Cyclin D1 provides a link between development and oncogenesis in the retina and breast. Cell. 1995, 82: 621-630. 10.1016/0092-8674(95)90034-9.
Xu X, Wagner KU, Larson D, Weaver Z, Li C, Ried T, Hennighausen L, Wynshaw-Boris A, Deng CX: Conditional mutation of BRCA1 in mammary epithelial cells results in blunted ductal morphogenesis and tumour formation. Nat Genet. 1999, 22: 37-43. 10.1038/8743.
Stacey D: Cyclin D1 serves as a cell cycle regulatory switch in actively proliferating cells. Curr Opin Cell Biol. 2003, 15: 158-163. 10.1016/S0955-0674(03)00008-5.
Morris JR, Pangon L, Boutell C, Katagiri T, Keep NH, Solomen ES: Genetic analysis of BRCA1 ubiquitin ligase activity and its relationship to breast cancer susceptibility. Hum Mol Genet. 2006, 15: 599-606. 10.1093/hmg/ddi476.
About this article
Cite this article
Murray, M., Harkin, D. Regulation of cyclin D1by the BRCA1–BARD1 complex. Breast Cancer Res 10 (Suppl 2), P22 (2008). https://doi.org/10.1186/bcr1906
- Breast Cancer
- Breast Cancer Development
- Ring Domain
- Polyubiquitin Chain