CARM1, coactivator associated arginine (R) methyltransferase 1, is involved in the activation of a number of transcriptional factors, including NF-?B, p53, E2F1 and steroid receptors, among which transcriptional activation of estrogen receptors (ERs) by CARM1 is best characterized. CARM1 methylates histone H3 at R17, and this methylation correlates with activation of the ER-target gene pS2. Loss of CARM1 leads to abrogation of the estrogen response and reduction in expression of some ER-target genes. We recently identified a CARM1-associated complex, nucleosomal methylation activator complex (NuMAC), which includes multiple components of SWI/SNF chromatin remodeling complex. SWI/SNF has also been implicated in ER-mediated transcriptional activation, and loss of SWI/SNF function is common in cancer progression. Interestingly, we have recently found that CARM1 can be phosphorylated in vivo and that its phosphorylation inhibits its histone methyltransferase (HMT) activity. We have generated phosphorylation-defective mutants of CARM1, which provide us a powerful tool to investigate the molecular mechanism of the regulation of ER-signaling by the CARM1 complex. This study is proposed to further understand the mode of transcriptional regulation and the cellular signaling of CARM1 in the context of ER. Given the surge of interest in histone arginine methylation in transcriptional regulation and the incorporation of two enzymatic activities, HMT and ATP-dependent remodeling, in the ER coactivator complex, our long-term objectives are to analyze the molecular underpinnings of ER transcriptional regulation by the CARM1 complex and to determine its functional relevance to the development of breast cancer. Our central hypothesis is that CARM1 and its associated enzymatic activities are important in regulating a subset of ER-target genes. CARM1 plays a central role in signal transduction, which is regulated by upstream cellular pathways and encodes a methyl-mark on histones to lead to the downstream transcription activation. Here I outline a series of experiments using biochemical, cell-based, and genomic approaches to study: (a) the molecular mechanism of the regulation of ER-target gene expression by CARM1 and SWI/SNF;(b) the effect of CARM1 phosphorylation on ER-dependent transcription and upstream signaling pathways and that lead to CARM1 inactivation;and (c) the downstream cellular effects of estrogen-dependent histone arginine methylation. These studies will provide a new understanding of the mechanism and functional significance of histone arginine methylation in ER-regulated processes.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA125387-05
Application #
8208236
Study Section
Molecular and Cellular Endocrinology Study Section (MCE)
Program Officer
Sathyamoorthy, Neeraja
Project Start
2008-04-01
Project End
2013-07-31
Budget Start
2012-02-01
Budget End
2013-07-31
Support Year
5
Fiscal Year
2012
Total Cost
$295,769
Indirect Cost
$94,494
Name
University of Wisconsin Madison
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Wang, Lu; Zhao, Zibo; Meyer, Mark B et al. (2014) CARM1 methylates chromatin remodeling factor BAF155 to enhance tumor progression and metastasis. Cancer Cell 25:21-36
Zhao, Zibo; Wang, Lu; Wen, Zhi et al. (2013) Systematic analyses of the cytotoxic effects of compound 11a, a putative synthetic agonist of photoreceptor-specific nuclear receptor (PNR), in cancer cell lines. PLoS One 8:e75198
Sievers, Chelsie K; Shanle, Erin K; Bradfield, Christopher A et al. (2013) Differential action of monohydroxylated polycyclic aromatic hydrocarbons with estrogen receptors ? and ?. Toxicol Sci 132:359-67
Wang, Lu; Charoensuksai, Purin; Watson, Nikole J et al. (2013) CARM1 automethylation is controlled at the level of alternative splicing. Nucleic Acids Res 41:6870-80
Shanle, Erin K; Zhao, Zibo; Hawse, John et al. (2013) Research resource: global identification of estrogen receptor * target genes in triple negative breast cancer cells. Mol Endocrinol 27:1762-75
Wen, Zhi; Pyeon, Dohun; Wang, Yidan et al. (2012) Orphan nuclear receptor PNR/NR2E3 stimulates p53 functions by enhancing p53 acetylation. Mol Cell Biol 32:26-35
Wu, Jiacai; Xu, Wei (2012) Histone H3R17me2a mark recruits human RNA polymerase-associated factor 1 complex to activate transcription. Proc Natl Acad Sci U S A 109:5675-80
Powell, Emily; Shanle, Erin; Brinkman, Ashley et al. (2012) Identification of estrogen receptor dimer selective ligands reveals growth-inhibitory effects on cells that co-express ER? and ER?. PLoS One 7:e30993
Chumanov, Robert S; Kuhn, Peter A; Xu, Wei et al. (2011) Expression and purification of full-length mouse CARM1 from transiently transfected HEK293T cells using HaloTag technology. Protein Expr Purif 76:145-53
Al-Dhaheri, Mariam; Wu, Jiacai; Skliris, Georgios P et al. (2011) CARM1 is an important determinant of ERýý-dependent breast cancer cell differentiation and proliferation in breast cancer cells. Cancer Res 71:2118-28

Showing the most recent 10 out of 19 publications