Data availabilityThe raw ChIP-seq, RNA-seq, CUT&Tag-seq and TT-seq data were deposited to the National Genomics Data Center under accession number HRA007983. All data supporting the findings of this study are available within the manuscript and its Supplementary Information. Source data are provided with this paper.ReferencesSabari, B. R., Zhang, D., Allis, C. D. & Zhao, Y. Metabolic regulation of gene expression through histone acylations. Nat. Rev. Mol. Cell Biol. 18, 90–101 (2017).Article CAS PubMed Google Scholar Wang, M. & Lin, H. Understanding the function of mammalian sirtuins and protein lysine acylation. Annu. Rev. Biochem. 90, 245–285 (2021).Article CAS PubMed Google Scholar Barnes, C. E., English, D. M. & Cowley, S. M. Acetylation & Co: an expanding repertoire of histone acylations regulates chromatin and transcription. Essays Biochem. 63, 97–107 (2019).Article CAS PubMed PubMed Central Google Scholar Tan, M. et al. 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We also thank members of the J.W. lab for valuable discussion.FundingThis study is supported by grants from the Ministry of Science and Technology of China (2023YFA1800402 to J.W. and W.W.), National Natural Science Foundation of China (32130051 and 32330020 to J.W., 82173073 to W.W.) and Fundamental Research Funds for the Central Universities (W.W.).Author informationAuthor notesThese authors contributed equally: Wei Wei, Zhongye Dai, Lei Shu.Authors and AffiliationsFengxian District Central Hospital-ECNU Joint Center of Translational Medicine, Institute of Biomedical Sciences and School of Life Sciences, Shanghai Key Laboratory of Regulatory Biology, East China Normal University, Shanghai, ChinaWei Wei, Zhongye Dai, Lei Shu, Meilin Sun, Jing Luo, Xixia Yuan, Song Gao, Xin Huang, Xiaotong Liu, Haonan Wen, Yunfeng Zhang, Yuanyong Huang, Wei Yang, Xianyu Huang, Lulu He, Siqi Lintong, Jiwen Li, Qiansen Zhang & Jiemin WongShanghai Diabetes Institute, Shanghai Key Laboratory of Diabetes Mellitus, Shanghai Clinical Centre for Diabetes, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, ChinaWei WeiDept. of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USADoug W. Chan, David M. Lonard, Jianming Xu & Bert W. O’MalleyState Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, ChinaBingbing Hao & Minjia TanDepartment of Biochemistry and Molecular Cell Biology, Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Shanghai Key Laboratory for Tumor Microenvironment and Inflammation, Shanghai Jiao Tong University School of Medicine, Shanghai, ChinaJingdong Xue & Bing LiDepartment of Urology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, ChinaJianjun YuState Key Laboratory of Genetic Engineering, School of Life Sciences and Zhongshan Hospital, Fudan University, Shanghai, ChinaGang WangJoint Center for Translational Medicine, Southern Medical University Affiliated Fengxian Hospital, Shanghai, ChinaZhenliang SunJoint Center for Translational Medicine, Shanghai Fifth People’s Hospital, Fudan University and School of Life Science, East China Normal University, Shanghai, ChinaJiemin WongAuthorsWei WeiView author publicationsSearch author on:PubMed Google ScholarZhongye DaiView author publicationsSearch author on:PubMed Google ScholarLei ShuView author publicationsSearch author on:PubMed Google ScholarMeilin SunView author publicationsSearch author on:PubMed Google ScholarDoug W. ChanView author publicationsSearch author on:PubMed Google ScholarBingbing HaoView author publicationsSearch author on:PubMed Google ScholarJingdong XueView author publicationsSearch author on:PubMed Google ScholarJing LuoView author publicationsSearch author on:PubMed Google ScholarXixia YuanView author publicationsSearch author on:PubMed Google ScholarSong GaoView author publicationsSearch author on:PubMed Google ScholarXin HuangView author publicationsSearch author on:PubMed Google ScholarXiaotong LiuView author publicationsSearch author on:PubMed Google ScholarHaonan WenView author publicationsSearch author on:PubMed Google ScholarYunfeng ZhangView author publicationsSearch author on:PubMed Google ScholarYuanyong HuangView author publicationsSearch author on:PubMed Google ScholarWei YangView author publicationsSearch author on:PubMed Google ScholarXianyu HuangView author publicationsSearch author on:PubMed Google ScholarLulu HeView author publicationsSearch author on:PubMed Google ScholarSiqi LintongView author publicationsSearch author on:PubMed Google ScholarJiwen LiView author publicationsSearch author on:PubMed Google ScholarQiansen ZhangView author publicationsSearch author on:PubMed Google ScholarJianjun YuView author publicationsSearch author on:PubMed Google ScholarGang WangView author publicationsSearch author on:PubMed Google ScholarDavid M. LonardView author publicationsSearch author on:PubMed Google ScholarJianming XuView author publicationsSearch author on:PubMed Google ScholarBing LiView author publicationsSearch author on:PubMed Google ScholarMinjia TanView author publicationsSearch author on:PubMed Google ScholarZhenliang SunView author publicationsSearch author on:PubMed Google ScholarBert W. O’MalleyView author publicationsSearch author on:PubMed Google ScholarJiemin WongView author publicationsSearch author on:PubMed Google ScholarContributionsW.W., Z.D., L.S., D.M.L., J. Xu, B.W.O. and J.W. conceptualized the project and designed the studies. W.W., Z.D. and J.W. wrote the manuscript, which was revised by D.M.L, J. Xu, B.W.O, J. Li and J.W. W.W., Z.D. and L.S. designed and performed most of the molecular and biochemistry experiments. X.L., Xianyu Huang, Y.H. and S.L.T. participated in the screening for HSTs in cells. D.W.C performed the in vitro affinity purification of Mediator proteins and their identification by LC–MS. W.W. and X.Y. performed the succinylated peptide pulldown experiments. G.W. provided plasmids encoding different subunits of the Mediator complex. Z.D., L.S., S.G., Xin Huang and L.H. generated NCOA3-KO, Ncoa3ki/ki and Ncoa1/2-DKO–Ncoa3–dTAG knock-in E14 cell lines and performed the RNA-seq, TT-seq, CUT&Tag and ChIP-seq experiments. W.W. and J. Luo performed the TT-seq and ChIP-seq of NCOA1/2/3-TKD SUM159PT cells. W.Y. purified the NCOA3(1102-C) and NCOA3(1102-C)-EL1232/3AA mutant proteins from E. coli. J. Xue and B.L. reconstructed the biotinylated mononucleosomes in vitro. M.S., Y.Z. and H.W. performed all bioinformatics data analysis. Q.Z. predicted the potential interaction between the AD2 region of NCOA-family proteins and succinyl-CoA using molecular docking. B.H. and M.T performed the MS analyses of histone succinylation. J.Y. and Z.S. provided critical reagents and contributed to data analysis.Corresponding authorsCorrespondence to Wei Wei or Jiemin Wong.Ethics declarationsCompeting interestsThe authors declare no competing interests.Peer reviewPeer review informationNature Chemical Biology thanks Kai Zhang and the other, anonymous reviewer(s) for their contribution to the peer review of this work. Peer reviewer reports are available.Additional informationPublisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.Extended dataExtended Data Fig. 1 Screening classic HATs for succinyltransferase activity (HST) by IF staining.a, IF staining screening of HST activity. HeLa cells were transfected with various HAT expression constructs as indicated. Two days after transfection, cells were processed for double IF staining using corresponding TAG-specific antibody and anti-pan-Ksu or pan-Kac antibody as indicated. Note for HAT1 IF staining was carried out using H4K5ac-specific antibody rather than pan-Kac antibody. Percentage represents the cells positive for elevated nuclear succinylation out of 30 counted transfected cells. Scale bar, 15 μm. b, Nuclear succinylation intensity was quantified using Image J by measuring mean nuclear fluorescence intensity in various HATs transfected cells. 30 cells were counted for each IF staining sample. Data are presented as mean ± SD from three independent biological replicates, **** P