Epigenetic regulation of the Warburg effect by H2B monoubiquitination


Cancer cells reprogram their energy metabolic system from the mitochondrial oxidative phosphorylation (OXPHOS) pathway to a glucose-dependent aerobic glycolysis pathway. This metabolic reprogramming phenomenon is known as the Warburg effect, a significant hallmark of cancer. However, the detailed mechanisms underlying this event or triggering this reprogramming remain largely unclear. Here, we found that histone H2B monoubiquitination (H2Bub1) negatively regulates the Warburg effect and tumorigenesis in human lung cancer cells (H1299 and A549 cell lines) likely through controlling the expression of multiple mitochondrial respiratory genes, which are essential for OXPHOS. Moreover, our work also suggested that pyruvate kinase M2 (PKM2), the rate-limiting enzyme of glycolysis, can directly interact with H2B in vivo and in vitro and negatively regulate the level of H2Bub1. The inhibition of cell proliferation and nude mice xenograft of human lung cancer cells induced by PKM2 knockdown can be partially rescued through lowering H2Bub1 levels, which indicates that the oncogenic function of PKM2 is achieved, at least partially, through the control of H2Bub1. Furthermore, PKM2 and H2Bub1 levels are negatively correlated in cancer specimens. Therefore, these findings not only provide a novel mechanism triggering the Warburg effect that is mediated through an epigenetic pathway (H2Bub1) but also reveal a novel metabolic regulator (PKM2) for the epigenetic mark H2Bub1. Thus, the PKM2-H2Bub1 axis may become a promising cancer therapeutic target.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8


  1. 1.

    Kim J, Hake SB, Roeder RG. The human homolog of yeast BRE1 functions as a transcriptional coactivator through direct activator interactions. Mol Cell. 2005;20:759–70.

  2. 2.

    Kim J, Guermah M, McGinty RK, Lee JS, Tang Z, Milne TA, et al. RAD6-Mediated transcription-coupled H2B ubiquitylation directly stimulates H3K4 methylation in human cells. Cell. 2009;137:459–71.

  3. 3.

    Chen S, Li J, Wang DL, Sun FL. Histone H2B lysine 120 monoubiquitination is required for embryonic stem cell differentiation. Cell Res. 2012;22:1402–5.

  4. 4.

    Fuchs G, Shema E, Vesterman R, Kotler E, Wolchinsky Z, Wilder S, et al. RNF20 and USP44 regulate stem cell differentiation by modulating H2B monoubiquitylation. Mol Cell. 2012;46:662–73.

  5. 5.

    Karpiuk O, Najafova Z, Kramer F, Hennion M, Galonska C, König A, et al. The histone H2B monoubiquitination regulatory pathway is required for differentiation of multipotent stem cells. Mol Cell. 2012;46:705–13.

  6. 6.

    Chen S, Jing Y, Kang X, Yang L, Wang DL, Zhang W, et al. Histone H2B monoubiquitination is a critical epigenetic switch for the regulation of autophagy. Nucleic Acids Res. 2017;45:1144–58.

  7. 7.

    Huang X, Yang L, Cai FF, Wang Y, Chen P, Mi J, et al. Autophagy-related protein ATG5 regulates histone H2B mono-ubiquitylation by translational control of RNF20. J Genet Genomics. 2017;44:503–6.

  8. 8.

    Spolverini A, Fuchs G, Bublik DR, Oren M. let-7b and let-7c microRNAs promote histone H2B ubiquitylation and inhibit cell migration by targeting multiple components of the H2B deubiquitylation machinery. Oncogene. 2017;36:5819–28.

  9. 9.

    Zhang K, Wang J, Tong TR, Wu X, Nelson R, Yuan YC, et al. Loss of H2B monoubiquitination is associated with poor-differentiation and enhanced malignancy of lung adenocarcinoma. Int J Cancer. 2017;141:766–77.

  10. 10.

    Tarcic O, Granit RZ, Pateras IS, Masury H, Maly B, Zwang Y, et al. RNF20 and histone H2B ubiquitylation exert opposing effects in Basal-Like versus luminal breast cancer. Cell Death Differ. 2017;24:694–704.

  11. 11.

    Tarcic O, Pateras IS, Cooks T, Shema E, Kanterman J, Ashkenazi H, et al. RNF20 links histone H2B ubiquitylation with inflammation and inflammation-associated cancer. Cell Rep. 2016;14:1462–76.

  12. 12.

    Prenzel T, Begus-Nahrmann Y, Kramer F, Hennion M, Hsu C, Gorsler T, et al. Estrogen-dependent gene transcription in human breast cancer cells relies upon proteasome-dependent monoubiquitination of histone H2B. Cancer Res. 2011;71:5739–53.

  13. 13.

    Liberti MV, Locasale JW. The Warburg effect: how doesit benefit cancer cells? Trends Biochem Sci. 2016;41:211–8.

  14. 14.

    Yang W, Xia Y, Hawke D, Li X, Liang J, Xing D, et al. PKM2 phosphorylates histone H3 and promotes gene transcription and tumorigenesis. Cell. 2012;150:685–96.

  15. 15.

    Shestov AA, Liu X, Ser Z, Cluntun AA, Hung YP, Huang L, et al. Quantitative determinants of aerobic glycolysis identify flux through the enzyme GAPDH as a limiting step. Elife. 2014;3:e03342.

  16. 16.

    Vander Heiden MG, Cantley LC, Thompson CB. Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science. 2009;324:1029–33.

  17. 17.

    DeBerardinis RJ, Lum JJ, Hatzivassiliou G, Thompson CB. The biology of cancer: metabolic reprogramming fuels cell growth and proliferation. Cell Metab. 2008;7:11–20.

  18. 18.

    Estrella V, Chen T, Lloyd M, Wojtkowiak J, Cornnell HH, Ibrahim-Hashim A, et al. Acidity generated by the tumor microenvironment drives local invasion. Cancer Res. 2013;73:1524–35.

  19. 19.

    Colegio OR, Chu NQ, Szabo AL, Chu T, Rhebergen AM, Jairam V, et al. Functional polarization of tumour-associated macrophages by tumour-derived lactic acid. Nature. 2014;513:559–63.

  20. 20.

    Ying H, Kimmelman AC, Lyssiotis CA, Hua S, Chu GC, Fletcher-Sananikone E, et al. Oncogenic Kras maintains pancreatic tumors through regulation of anabolic glucose metabolism. Cell. 2012;149:656–70.

  21. 21.

    Luo W, Hu H, Chang R, Zhong J, Knabel M, O’Meally R, et al. Pyruvate kinase M2 is a PHD3-stimulated coactivator for hypoxia-inducible factor1. Cell. 2011;145:732–44.

  22. 22.

    Yang W, Xia Y, Ji H, Zheng Y, Liang J, Huang W, et al. Nuclear PKM2 regulates β-catenin transactivation upon EGFR activation. Nature. 2011;480:118–22.

  23. 23.

    Gao X, Wang H, Yang JJ, Liu X, Liu ZR. Pyruvate kinase M2 regulates gene transcription by acting as a protein kinase. Mol Cell. 2012;45:598–609.

  24. 24.

    Jiang Y, Li X, Yang W, Hawke DH, Zheng Y, Xia Y, et al. PKM2 regulates chromosome segregation and mitosis progression of tumor cells. Mol Cell. 2014;53:75–87.

  25. 25.

    Jiang Y, Wang Y, Wang T, Hawke DH, Zheng Y, Li X, et al. PKM2 phosphorylates MLC2 and regulates cytokinesis of tumour cells. Nat Commun. 2014;5:5566.

  26. 26.

    He CL, Bian YY, Xue Y, Liu ZX, Zhou KQ, Yao CF, et al. Pyruvate kinase M2 activates mTORC1 by phosphorylating AKT1S1. Sci Rep. 2016;6:21524.

  27. 27.

    Wei Y, Wang D, Jin F, Bian Z, Li L, Liang H, et al. Pyruvate kinase type M2 promotes tumour cell exosome release via phosphorylating synaptosome-associated protein 23. Nat Commun. 2017;8:14041.

  28. 28.

    Hosios AM, Fiske BP, Gui DY, Vander Heiden MG. Lack of evidence for PKM2 protein kinase activity. Mol Cell. 2015;59:850–7.

  29. 29.

    Riscal R, Schrepfer E, Arena G, Cissé MY, Bellvert F, Heuillet M, et al. Chromatin-bound MDM2 regulates serine metabolism and redox homeostasis independently of p53. Mol Cell. 2016;62:890–902.

  30. 30.

    Warburg O. On the origin of cancer cells. Science. 1956;123:309–14.

  31. 31.

    Fantin VR, St-Pierre J, Leder P. Attenuation of LDH-A expression uncovers a link between glycolysis, mitochondrialphysiology, and tumor maintenance. Cancer Cell. 2006;9:425–34.

  32. 32.

    Moreno-Sánchez R, Rodríguez-Enríquez S, Marín-Hernández A, Saavedra E. Energy metabolism in tumor cells. FEBS J. 2007;274:1393–418.

  33. 33.

    Bellance N, Benard G, Furt F, Begueret H, Smolková K, Passerieux E, et al. Bioenergetics of lung tumors: alteration of mitochondrial biogenesis and respiratory capacity. Int J Biochem Cell Biol. 2009;41:2566–77.

  34. 34.

    Jose C, Bellance N, Rossignol R. Choosing between glycolysis and oxidative phosphorylation: a tumor’s dilemma? Biochim Biophys Acta. 2011;1807:552–61.

  35. 35.

    Christofk HR, Vander Heiden MG, Harris MH, Ramanathan A, Gerszten RE, Wei R, et al. The M2 splice isoform of pyruvate kinase is important for cancer metabolism and tumour growth. Nature. 2008;452:230–3.

  36. 36.

    Mellati AA, Yücel M, Altinörs N, Gündüz U. Regulation of M2-type pyruvate kinase from human meningioma by allosteric effectors fructose 1,6 diphosphate and L-alanine. Cancer Biochem Biophys. 1992;13:33–41.

  37. 37.

    Chen Z, Wang Z, Guo W, Zhang Z, Zhao F, Zhao Y, et al. TRIM35 Interacts with pyruvate kinase isoform M2 to suppress the Warburg effect and tumorigenicity in hepatocellular carcinoma. Oncogene. 2015;34:3946–56.

  38. 38.

    Wu Y, Chen P, Jing Y, Wang C, Men YL, Zhan W, et al. Microarray analysis reveals potential biological functions of histone H2B monoubiquitination. PLoS One. 2015;10:e0133444.

  39. 39.

    Tekade RK, Sun X. The Warburg effect and glucose-derived cancer theranostics. Drug Disco Today. 2017;22:1637–53.

  40. 40.

    Christofk HR, Vander Heiden MG, Wu N, Asara JM, Cantley LC. Pyruvate kinase M2 is a phosphotyrosine-binding protein. Nature. 2008;452:181–6.

  41. 41.

    Anastasiou D, Poulogiannis G, Asara JM, Boxer MB, Jiang JK, Shen M, et al. Inhibition of pyruvate kinase M2 by reactive oxygen species contributes to cellular antioxidant responses. Science. 2011;334:1278–83.

  42. 42.

    Yang W, Zheng Y, Xia Y, Ji H, Chen X, Guo F, et al. ERK1/2-dependent phosphorylation and nuclear translocation of PKM2 promotes the Warburg effect. Nat Cell Biol. 2012;14:1295–304.

  43. 43.

    Li S, Swanson SK, Gogol M, Florens L, Washburn MP, Workman JL, et al. Serine and SAM responsive complex SESAME regulates histone modification crosstalk by sensing cellular metabolism. Mol Cell. 2015;60:408–21.

  44. 44.

    Hitosugi T, Kang S, Vander Heiden MG, Chung TW, Elf S, Lythgoe K, et al. Tyrosine phosphorylation inhibits PKM2 to promote the Warburg effect and tumor growth. Sci Signal. 2009;2:ra73.

  45. 45.

    Liang J, Cao R, Zhang Y, Xia Y, Zheng Y, Li X, et al. PKM2 dephosphorylation by Cdc25A promotes the Warburg effect and tumorigenesis. Nat Commun. 2016;7:12431.

  46. 46.

    Chao TK, Huang TS, Liao YP, Huang RL, Su PH, Shen HY, et al. Pyruvate kinase M2 is a poor prognostic marker of and a therapeutic target in ovarian cancer. PLoS One. 2017;12:e0182166.

  47. 47.

    Wang C, Jiang J, Ji J, Cai Q, Chen X, Yu Y, et al. PKM2 promotes cell migration and inhibits autophagy by mediating PI3K/AKT activation and contributes to the malignant development of gastric cancer. Sci Rep. 2017;7:2886.

  48. 48.

    Smolková K, Plecitá-Hlavatá L, Bellance N, Benard G, Rossignol R, Ježek P. Waves of gene regulation suppress and then restore oxidative phosphorylation in cancer cells. Int J Biochem Cell Biol. 2011;43:950–68.

  49. 49.

    Chen S, Wang DL, Liu Y, Zhao L, Sun FL. RAD6 regulates the dosage of p53 by a combination of transcriptional and posttranscriptional mechanisms. Mol Cell Biol. 2012;32:576–87.

  50. 50.

    Chen S, Wei HM, Lv WW, Wang DL, Sun FL. E2 ligase dRad6 regulates DMP53 turnover in Drosophila. J Biol Chem. 2011;286:9020–30.

  51. 51.

    An H, Yang L, Wang C, Gan Z, Gu H, Zhang T. et al. Interactome analysis reveals a novel role for RAD6 in the regulation of proteasome activity and localization in response to DNA damage. Mol Cell Biol. 2017;37:e00419–16.

Download references


We thank Dr Weiwei Yang from the Shanghai Institute of Biochemistry and Cell Biology (SIBS, CAS) for kindly providing the PKM2-related plasmids. This work was supported by the National Key Research and Development Program of China (Grant No.: 2017YFA0103301 and 2016YFA0100403), the 973 program of the Ministry of Science and Technology of China (Grant No.: 2015CB856204, 2015CB964802), the National Natural Science Foundation of China (Grant No.: 91419304, 81773009, 81972650, 31330043, and 31271534), the Fundamental Research Funds for the Central Universities (Xi’an Jiao Tong University, Grant No.: 2017qngz13), and the China Postdoctoral Science Foundation (Grant No.: 2017M613149 and 2018T111038).

Author information

Correspondence to Xiao-Mei Yang or Fang-Lin Sun or Su Chen.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Edited by M. Piacentini

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Jing, Y., Cai, F., Zhang, L. et al. Epigenetic regulation of the Warburg effect by H2B monoubiquitination. Cell Death Differ (2019). https://doi.org/10.1038/s41418-019-0450-2

Download citation