Position title: Helmut Beinert Professor of Biological Sciences
Phone: Phone: (608) 263-3975
RESEARCH INTERESTS - Physiological Mechanisms Associated with P450 Cytochromes. Our research addresses signaling mechanisms involving P450 cytochromes. These regulate activity at nuclear receptors regulated by three types of activator. - Cholesterol/steroid hormones - Fatty acids and metabolites - Ah receptor/PCB's and polycyclic aromatic hydrocarbons
BA/BS, Oxford University
Ph.D., Oxford University
Postdoctoral Research, Basel University, Switzerland
Project A: Regulation of StAR
We are examining regulation of cholesterol conversion to steriods that activate nuclear receptors. This requires cholesterol metabolism at P450 sec that is located in the mitochondria of specialized cells. This depends on the StAR protein which is regulated by cAMP at the level of transcription, mRNA stability and protein modification. Our recent work implicates additional regulation by various MAP kinases that regulate StAR mRNA stability via specific elements at the 3′ end of the mRNA. We are engineering the StAR mRNA to alter the protein function and mRNA stabiltity
Project B: Ah receptor suppresses differentiation via MAP kinases
Here we are dissecting the mechanism of differentiation of multipotential embryo cells to adipcytes. This process is activated by the PPAR(Gamma) receptor. We have shown that activation of the Ah receptor blocks adipogenesis by suppressing PPAR(Gamma) expression. This depends on activation of MAP kinases. The Ah receptor also regulates a cytochrome P4+50 in these cells (CCYP1B1) which may metabolize an endogenous activator of differentiation.
Project C: Cytochrome P4501B1 is regulated by both AhR and cAMP
We are studying the molecular regulation of CYP1B1 expression and the role this protein plays in cancer caused by polycyclic aromatic hydrocarbons. Using a CYP1B1 – knockout mouse we showed that metabolism by CYP1B1 is essential for many cancers caused by these chemicals. We have identified two enhancers regions in the CYP1B1 gene: one is very responsive to the Ah receptor, one responds to cAMP. There mechanisms are being further characterized in multiple cell types including mammary epithelia that are targets for this carcinogenesis. We are looking for determinants of cell specific expression and also testing the hypothessis that the key role of CYP1B1 in carcinogenesis arises from expression in susceptible multi-potential cells.
The laboratory studies the toxicity of various agents by identifying molecular mechanisms by which these processes are disrupted. The toxicity of sodium arsenite, TCDD (Dioxin) and polycyclic hydrocarbons is being studied in Projects A, B and C respectively.
- Lee J, Jefcoate C. (2017) Monitoring of Dual CRISPR/Cas9-Mediated Steroidogenic Acute Regulatory Protein Gene Deletion and Cholesterol Accumulation Using High-Resolution Fluorescence In Situ Hybridization in a Single Cell. Front Endocrinol (Lausanne). 2017 Oct 25;8:289.
- Lee J, Yamazaki T, Dong H, Jefcoate C.(2017) A single cell level measurement of StAR expression and activity in adrenal cells. Mol Cell Endocrinol. 2017 Feb 5;441:22-30. Epub 2016 Aug 10. Review.
- Maguire M, Larsen MC, Foong YH, Tanumihardjo S, Jefcoate CR. (2017) Cyp1b1 deletion and retinol deficiency coordinately suppress mouse liver lipogenic genes and hepcidin expression during post-natal development. Mol Cell Endocrinol. 2017 Oct 15;454:50-68.
- Maguire M, Bushkofsky JR, Larsen MC, Foong YH, Tanumihardjo SA, Jefcoate CR. (2017) Diet-dependent retinoid effects on liver gene expression include stellate and inflammation markers and parallel effects of the nuclear repressor Shp. J Nutr Biochem. Sep;47:63-74.
- Larsen MC, N’Jai AU, Alexander DL, Rondelli CM, Forsberg EC, Czuprynski CJ, Jefcoate CR. (2016) Cyp1b1-mediated suppression of lymphoid progenitors in bone marrow by polycyclic aromatic hydrocarbons coordinately impacts spleen and thymus: a selective role for the Ah Receptor. Pharmacol Res Perspect. Jul 29;4(4):e00245.
- Rondelli CM, Larsen MC, N’jai A, Czuprynski CJ, Jefcoate CR. PAHs Target Hematopoietic Linages in Bone Marrow through Cyp1b1 Primarily in Mesenchymal Stromal Cells but Not AhR: A Reconstituted In Vitro Model. Stem Cells Int. 2016;2016:1753491.
- Lee J, Tong T, Duan H, Foong YH, Musaitif I, Yamazaki T, Jefcoate C. (2016) Regulation of StAR by the N-terminal Domain and Coinduction of SIK1 and TIS11b/Znf36l1 in Single Cells. Front Endocrinol (Lausanne). 7:107.
- Lee J, Foong YH, Musaitif I, Tong T, Jefcoate C. (2016) Analysis of specific RNA in cultured cells through quantitative integration of q-PCR and N-SIM single cell FISH images: Application to hormonal stimulation of StAR transcription. Mol Cell Endocrinol. 5;429:93-105.
- Kim SH, Trinh AT, Larsen MC, Mastrocola AS, Jefcoate CR, Bushel PR, Tibbetts RS. (2016) Tunable regulation of CREB DNA binding activity couples genotoxic stress response and metabolism. Nucleic Acids Res. Jul 18. pii: gkw643. [Epub ahead of print]
- Dinu D, Chu C, Veith A, Lingappan K, Couroucli X, Jefcoate CR, Sheibani N, Moorthy B. (2016) Mechanistic role of cytochrome P450 (CYP)1B1 in oxygen-mediated toxicity in pulmonary cells: A novel target for prevention of hyperoxic lung injury. Biochem Biophys Res Commun. 476:346-51
- Bushkofsky JR, Maguire M, Larsen MC, Fong YH, Jefcoate CR. (2016) Cyp1b1 affects external control of mouse hepatocytes, fatty acid homeostasis and signaling involving HNF4α and PPARα. Arch Biochem Biophys. 597:30-47.
- Lee J, Tong T, Takemori H, Jefcoate C. (2015) Stimulation of StAR expression by cAMP is controlled by inhibition of highly inducible SIK1 via CRTC2, a co-activator of CREB. Mol Cell Endocrinol. 408:80-9.
- Larsen MC, Bushkofsky JR, Gorman T, Adhami V, Mukhtar H, Wang S, Reeder SB,
Sheibani N, Jefcoate CR. (2015) Cytochrome P450 1B1: An unexpected modulator of liver fatty acid homeostasis. Arch Biochem Biophys. 571:21-39.
- Li F, Jiang C, Larsen MC, Bushkofsky J, Krausz KW, Wang T, Jefcoate CR, Gonzalez FJ. (2014) Lipidomics reveals a link between CYP1B1 and SCD1 in promoting obesity. J Proteome Res. 13:2679-87.
- Piccinato CA, Rosa GJ, N’jai AU, Jefcoate CR, Wiltbank MC. (2013) Estradiol and progesterone exhibit similar patterns of hepatic gene expression regulation in the bovine model. PLoS One. 8:e73552.
- Zhao Y, Wang S, Sorenson CM, Teixeira L, Dubielzig RR, Peters DM, Conway SJ,
Jefcoate CR, Sheibani N. (2013) Cyp1b1 mediates periostin regulation of trabecular
meshwork development by suppression of oxidative stress. Mol Cell Biol. 33:4225-40.
- Palenski TL, Sorenson CM, Jefcoate CR, Sheibani N. (2013) Lack of Cyp1b1 promotes the proliferative and migratory phenotype of perivascular supporting cells. Lab Invest. 93:646-62.
- Zheng W, Tong T, Lee J, Liu X, Marcus C, Jefcoate CR. (2013) Stimulation of mouse Cyp1b1 during adipogenesis: characterization of promoter activation by the transcription factor Pax6. Arch Biochem Biophys. 532:1-14.
- N’jai AU, Larsen MC, Bushkofsky JR, Czuprynski CJ, Jefcoate CR. (2011) Acute disruption of bone marrow hematopoiesis by benzo(a)pyrene is selectively reversed by aryl hydrocarbon receptor-mediated processes. Mol Pharmacol. 79:724-34.
- Jefcoate CR, Lee J, Cherradi N, Takemori H, Duan H. (2011) cAMP stimulation of StAR expression and cholesterol metabolism is modulated by co-expression of labile suppressors of transcription and mRNA turnover. Mol Cell Endocrinol. 336:53-62.
- N’jai AU, Larsen M, Shi L, Jefcoate CR, Czuprynski CJ. (2010) Bone marrow lymphoid and myeloid progenitor cells are suppressed in 7,12-dimethylbenz(a)anthracene (DMBA) treated mice. Toxicology. 271:27-35.
- Tang Y, Scheef EA, Gurel Z, Sorenson CM, Jefcoate CR, Sheibani N.(2010) CYP1B1 and endothelial nitric oxide synthase combine to sustain proangiogenic functions of endothelial cells under hyperoxic stress. Am J Physiol Cell Physiol. 298:C665-78.
- Tang Y, Scheef EA, Wang S, Sorenson CM, Marcus CB, Jefcoate CR, Sheibani N. (2009) CYP1B1 expression promotes the proangiogenic phenotype of endothelium through decreased intracellular oxidative stress and thrombospondin-2 expression. Blood. 113:744-54.
- Halberg RB, Larsen MC, Elmergreen TL, Ko AY, Irving AA, Clipson L, Jefcoate
CR. (2008) Cyp1b1 exerts opposing effects on intestinal tumorigenesis via exogenous and endogenous substrates. Cancer Res. 68:7394-402.
- Jefcoate CR, Wang S, Liu X. (2008) Methods that resolve different contributions of
clonal expansion to adipogenesis in 3T3-L1 and C3H10T1/2 cells. Methods Mol Biol. 456:173-93.
- Duan H and Jefcoate CR. (2007). The predominant cAMP-stimulated 3 x 5 kb StAR mRNA contains specific sequence elements in the extended 3’UTR that confer high basal instability. J Mol Endocrinol. 38:159-179. PMID 17242178
- Jefcoate CR. (2006). Liver X receptor opens a new gateway to StAR and to steroid hormones. J Clin Invest. 116:1832-5. PMID 16823483
- Galvan N, Page TJ, Czuprynski CJ, Jefcoate CR. (2006). Benzo(a)pyrene and 7,12-dimethylbenz(a)anthrecene differentially affect bone marrow cells of the lymphoid and myeloid lineages. Toxicol Appl Pharmacol. 213:105-16. PMID 16307768
- Hanlon PR, Cimafranca MA, Liu X, Cho YC, Jefcoate CR. (2005). Microarray analysis of early adipogenesis in C3H10T1/2 cells: cooperative inhibitory effects of growth factors and 2,3,7,8-tetrachlorodibenzo-p-dioxin. Toxicol Appl Pharmacol. 207:39-58. PMID 16054899
- Zhao D, Duan H, Kim YC, Jefcoate CR. (2005). Rodent StAR mRNA is substantially regulated by control of mRNA stability through sites in the 3′-untranslated region and through coupling to ongoing transcription. J Steroid Biochem Mol Biol. 96:155-73. PMID 16039847
- Yuroff AS, Jefcoate CR, Czuprynski CJ. (2005). Close proximity, but not VLA-4-dependent adherence between pre-B cells and bone marrow stromal cells, is required for DMBA-induced apoptosis of pre-B cells in vitro. Toxicol Lett. 156:253-60. PMID 15737488
- Galvan N, Teske DE, Zhou G, Moorthy B, MacWilliams PS, Czuprynski CJ, Jefcoate CR. (2005). Induction of CYP1A1 and CYP1B1 in liver and lung by benzo(a)pyrene and 7,12-d imethylbenz(a)anthracene do not affect distribution of polycyclic hydrocarbons to target tissue: role of AhR and CYP1B1 in bone marrow cytotoxicity. Toxicol Appl Pharmacol. 202:244-57. PMID 15667830
- Hanlon PR, Zheng W, Ko AY, Jefcoate CR. (2005). Identification of novel TCDD-regulated genes by microarray analysis. Toxicol Appl Pharmacol. 202:215-28. PMID 15667827
- Cho YC, Zheng W, Yamamoto M, Liu X, Hanlon PR, and Jefcoate CR. (2005). Differentiation of pluripotent C3H10T1/2 cells rapidly elevates CYP1B1 through a novel process that overcomes a loss of Ah Receptor. Arch Biochem Biophys. 439:139-153. PMID 15967407
- Larsen MC, Brake PB, Pollenz RS, and Jefcoate CR. (2004). Linked expression of Ah receptor, ARNT, CYP1A1, and CYP1B1 in rat mammary epithelia, in vitro, is each substantially elevated by specific extracellular matrix interactions that precede branching morphogenesis. Toxicol Sci. 82:46-61. PMID 15297627
- Cho YC, Zheng W, and Jefcoate CR. (2004). Disruption of cell-cell contact maximally but transiently activates AhR-mediated transcription in 10T1/2 fibroblasts. Toxicol Appl Pharmacol. 199:220-238. PMID 15364539
- Page TJ, MacWilliams PS, Suresh M, Jefcoate CR, and Czuprynski CJ. (2004). 7-12 Dimethylbenz[a]anthracene-induced bone marrow hypocellularity is dependent on signaling through both the TNFR and PKR. Toxicol Appl Pharmacol. 198:21-28. PMID 15207645