caa a22 4500 60623389X CHVBK 20210128101234.0 cr unu---uuuuu 210128e20150401xx s 000 0 eng 10.1007/s11010-014-2310-1 doi (NATIONALLICENCE)springer-10.1007/s11010-014-2310-1 DNA methylation regulates bromodomain-containing protein 2 expression during adipocyte differentiation [Elektronische Daten] [Ruixin Sun, Yi Wu, Yuxiong Wang, Kun Zang, Huanhuan Wei, Fangnian Wang, Min Yu] Obesity is characterized by excessive accumulation of white adipose tissue. Bromodomain-containing protein 2 (Brd2), which belongs to the bromodomain and extraterminal domain family of proteins, suppresses adipocyte differentiation. DNA methylation is critical for several differentiation processes and possibly in adipocyte differentiation. However, whether DNA methylation regulates the expression of Brd2 is not clear. In our study, we demonstrated that DNA methylation contributes to the regulation of Brd2 expression during pre-adipocyte differentiation. Brd2 mRNA levels were low in pre-adipocytes, increased in early adipocytes, and declined in mature adipocytes. To test whether and how Brd2 expression is regulated by DNA methylation during the differentiation of 3T3-L1 pre-adipocytes to adipocytes, cells were cultured in the presence of the methylation inhibitor 5-aza-2′-deoxycytidine (5-Aza). Pre-adipocytes and adipocytes exposed to 5-Aza exhibited a dose-dependent increase in Brd2 transcription levels, while only mature adipocytes exhibited increased expression of Brd2 protein. Subsequently, we tested the DNA methylation status of the Brd2 promoter region. Bisulfite-sequencing analysis revealed that six CpG sites in two predicted promoters of Brd2 were demethylated in early adipocytes and highly methylated in mature adipocytes. Digestion of bisulfite-converted PCR products of the Brd2 promoter region from 3T3-L1 cells with BstU1 (CGGC) revealed that the demethylation rate of the Brd2 promoter was consistent with Brd2 mRNA expression in differentiating 3T3-L1 cells. In conclusion, DNA demethylation of the Brd2 promoter region induced Brd2 expression during differentiation of 3T3-L1 cells into adipocytes. Springer Science+Business Media New York, 2015 Brd2 nationallicence DNA methylation nationallicence Bisulfite-sequencing analysis nationallicence Adipocyte differentiation nationallicence Sun Ruixin The Key Laboratory of Molecular Medicine, The Ministry of Education, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, No. 138 Yi Xue Yuan Road, P.O. Box #238, 200032, Shanghai, China aut Wu Yi The Key Laboratory of Molecular Medicine, The Ministry of Education, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, No. 138 Yi Xue Yuan Road, P.O. Box #238, 200032, Shanghai, China aut Wang Yuxiong The Key Laboratory of Molecular Medicine, The Ministry of Education, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, No. 138 Yi Xue Yuan Road, P.O. Box #238, 200032, Shanghai, China aut Zang Kun The Key Laboratory of Molecular Medicine, The Ministry of Education, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, No. 138 Yi Xue Yuan Road, P.O. Box #238, 200032, Shanghai, China aut Wei Huanhuan The Key Laboratory of Molecular Medicine, The Ministry of Education, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, No. 138 Yi Xue Yuan Road, P.O. Box #238, 200032, Shanghai, China aut Wang Fangnian The Key Laboratory of Molecular Medicine, The Ministry of Education, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, No. 138 Yi Xue Yuan Road, P.O. Box #238, 200032, Shanghai, China aut Yu Min The Key Laboratory of Molecular Medicine, The Ministry of Education, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, No. 138 Yi Xue Yuan Road, P.O. Box #238, 200032, Shanghai, China aut Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 402/1-2(2015-04-01), 23-31 0300-8177 402:1-2<23 2015 402 11010 https://doi.org/10.1007/s11010-014-2310-1 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s11010-014-2310-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sun Ruixin The Key Laboratory of Molecular Medicine, The Ministry of Education, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, No. 138 Yi Xue Yuan Road, P.O. Box #238, 200032, Shanghai, China aut NATIONALLICENCE 700 1- Wu Yi The Key Laboratory of Molecular Medicine, The Ministry of Education, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, No. 138 Yi Xue Yuan Road, P.O. Box #238, 200032, Shanghai, China aut NATIONALLICENCE 700 1- Wang Yuxiong The Key Laboratory of Molecular Medicine, The Ministry of Education, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, No. 138 Yi Xue Yuan Road, P.O. Box #238, 200032, Shanghai, China aut NATIONALLICENCE 700 1- Zang Kun The Key Laboratory of Molecular Medicine, The Ministry of Education, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, No. 138 Yi Xue Yuan Road, P.O. Box #238, 200032, Shanghai, China aut NATIONALLICENCE 700 1- Wei Huanhuan The Key Laboratory of Molecular Medicine, The Ministry of Education, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, No. 138 Yi Xue Yuan Road, P.O. Box #238, 200032, Shanghai, China aut NATIONALLICENCE 700 1- Wang Fangnian The Key Laboratory of Molecular Medicine, The Ministry of Education, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, No. 138 Yi Xue Yuan Road, P.O. Box #238, 200032, Shanghai, China aut NATIONALLICENCE 700 1- Yu Min The Key Laboratory of Molecular Medicine, The Ministry of Education, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, No. 138 Yi Xue Yuan Road, P.O. Box #238, 200032, Shanghai, China aut NATIONALLICENCE 773 0- Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 402/1-2(2015-04-01), 23-31 0300-8177 402:1-2<23 2015 402 11010