Genetic Incorporation of N(ε)-Formyllysine, a New Histone Post-translational Modification.
Lysine formylation is a newly discovered post-translational modification (PTM) in histones and other nuclear proteins; it has a well-recognized but poorly defined role in chromatin conformation modulation and gene expression. To date, there is no general method to site-specifically incorporate N(ε)-formyllysine at a defined site of a protein. Here we report the highly efficient genetic incorporation of the unnatural amino acid N(ε)-formyllysine into proteins produced in Escherichia coli and mammalian cells, by using an orthogonal N(ε)-formyllysine tRNAsynthetase/tRNACUA pair. This technique can be applied to study the role of lysine formylation in epigenetic regulation.
School of Earth and Space Science, University of Science and Technology of China (USTC), 96 Jinzhai Road, Hefei, Anhui 230026 (China).
Tianyuan
T
Wang
Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101 (China). zqboy008@163.com.
Qing
Q
Zhou
Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101 (China).
Fahui
F
Li
Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin 300308 (China).
Yang
Y
Yu
School of Earth and Space Science, University of Science and Technology of China (USTC), 96 Jinzhai Road, Hefei, Anhui 230026 (China). xbyin@ustc.edu.cn.|Suzhou Key Lab for Selenium and Human Health, Suzhou Institute for Advanced Study, USTC, 166 Ren'ai Road, Dushu Lake Higher Education Area, Industrial Park, Suzhou, Jiangsu 215123 (China). xbyin@ustc.edu.cn.
Xuebin
X
Yin
Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101 (China). jwang@ibp.ac.cn.
Jiangyun
J
Wang
D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D004926:Escherichia coli / Q000235:genetics; D006657:Histones / Q000235:genetics*; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives* / Q000235:genetics; D017021:Methanosarcina barkeri / Q000201:enzymology / Q000235:genetics; D011499:Protein Processing, Post-Translational
D006657:Histones; C020334:N(epsilon)-formyllysine; D000604:Amino Acyl-tRNA Synthetases; D008239:Lysine
amino acids; formyllysine; gene technology; histones; post-translational modifications