Rnf20 deficiency in adipocyte impairs adipose tissue development and thermogenesis

Abstract

RNF20, an E3 ligase critical for monoubiquitination of histone H2B at lysine 120 (H2Bub), has been implicated in the regulation of various cellar processes; however, its physiological roles in adipocytes remain poorly characterized. Here, we report that the adipocyte-specific knockout of Rnf20 (ASKO) in mice led to progressive fat loss, organomegaly and hyperinsulinemia. Despite signs of hyperinsulinemia, normal insulin sensitivity and improved glucose tolerance were observed in the young and aged CD-fed ASKO mice. In addition, high-fat dietfed ASKO mice developed severe liver steatosis. Moreover, we observed that the ASKO mice were extremely sensitive to a cold environment due to decreased expression levels of brown adipose tissue (BAT) selective genes, including uncoupling protein 1 (Ucp1), and impaired mitochondrial functions. Significantly decreased levels of peroxisome proliferator-activated receptor gamma (Pparγ) were observed in the gonadal white adipose tissues (gWAT) from the ASKO mice, suggesting that Rnf20 regulates adipogenesis, at least in part, through Pparγ. Rosiglitazone-treated ASKO mice exhibited increased fat mass compared to that of the non-treated ASKO mice. Collectively, our results illustrate the critical role of RNF20 in control of white and brown adipose tissue development and physiological function.

Keywords

RNF20 / fat loss / adipose tissue development / thermogenesis

References

[1] Arioglu E, Duncan-Morin J, Sebring N, Rother KI, Gottlieb N, Lieberman J, Herion D, Kleiner DE, Reynolds J, Premkumar A, (2000) Efficacy and safety of troglitazone in the treatment of lipodystrophy syndromes. Ann Intern Med 133(4):263–274

[2] Boucher J,Mori MA, Lee KY, Smyth G, Liew CW, Macotela Y, Rourk M, Bluher M, Russell SJ, Kahn CR (2012) Impaired thermogenesis and adipose tissue development in mice with fat-specific disruption of insulin and IGF-1 signalling. Nat Commun 3:902

[3] Dawson MA, Kouzarides T, Huntly BJ (2012) Targeting epigenetic readers in cancer. N Engl J Med 367(7):647–657

[4] Duan Y, Huo D, Gao J, Wu H, Ye Z, Liu Z, Zhang K, Shan L, Zhou X, Wang Y, (2016) Ubiquitin ligase RNF20/40 facilitates spindle assembly and promotes breast carcinogenesis through stabilizing motor protein Eg5. Nat Commun 7:12648

[5] Duteil D, Tosic M, Lausecker F, Nenseth HZ, Muller JM, Urban S, Willmann D, Petroll K, Messaddeq N, Arrigoni L (2016) Lsd1 ablation triggers metabolic reprogramming of brown adipose tissue. Cell Rep 17(4):1008–1021

[6] Fierz B, Chatterjee C, Mcginty RK, Bar-Dagan M, Raleigh DP, Muir TW (2011) Histone H2B ubiquitylation disrupts local and higherorder chromatin compaction. Nat Chem Biol 7(2):113–119

[7] Fuchs G, Oren M (2014) Writing and reading H2B monoubiquitylation. Biochim Biophys Acta 1839(8):694–701

[8] Fuchs G, Shema E, Vesterman R, Kotler E, Wolchinsky Z, Wilder S, Golomb L, Pribluda A, Zhang F, Haj-Yahya M (2012) RNF20 and USP44 regulate stem cell differentiation by modulating H2B monoubiquitylation. Mol Cell 46(5):662–673

[9] Himms-Hagen J, Melnyk A, Zingaretti MC, Ceresi E, Barbatelli G, Cinti S (2000) Multilocular fat cells in WAT of CL-316243-treated rats derive directly from white adipocytes. Am J Physiol Cell Physiol 279(3):C670–681

[10] Jeon YG, Lee JH, Ji Y,Sohn JH, Lee D, Kim DW, Yoon SG, Shin KC, Park J, Seong JK (2020) RNF20 functions as a transcriptional coactivator for PPARgamma by promoting NCoR1 degradation in adipocytes. Diabetes 69(1):20–34

[11] Jernas M, Palming J, Sjoholm K, Jennische E, Svensson PA, Gabrielsson BG, Levin M, Sjogren A, Rudemo M, Lystig TC (2006) Separation of human adipocytes by size: hypertrophic fat cells display distinct gene expression. FASEB J 20(9):1540–1542

[12] Jin W, Patti ME (2009) Genetic determinants and molecular pathways in the pathogenesis of Type 2 diabetes. Clin Sci (Lond) 116(2):99–111

[13] Karpiuk O, Najafova Z, Kramer F, Hennion M, Galonska C, Konig A, Snaidero N, Vogel T, Shchebet A, Begus-Nahrmann Y (2012) The histone H2B monoubiquitination regulatory pathway is required for differentiation of multipotent stem cells. Mol Cell 46 (5):705–713

[14] Kim JI, Huh JY, Sohn JH, Choe SS, Lee YS, Lim CY, Jo A, Park SB, Han W, Kim JB (2015) Lipid-overloaded enlarged adipocytes provoke insulin resistance independent of inflammation. Mol Cell Biol 35(10):1686–1699

[15] Kouzarides T (2007) Chromatin modifications and their function. Cell 128(4):693–705

[16] Kubota N, Terauchi Y, Miki H, Tamemoto H, Yamauchi T, Komeda K, Satoh S, Nakano R, Ishii C, Sugiyama T (1999) PPAR gamma mediates high-fat diet-induced adipocyte hypertrophy and insulin resistance. Mol Cell 4(4):597–609

[17] Lee JH, Lee GY, Jang H, Choe SS, Koo SH, Kim JB (2014) Ring finger protein20 regulates hepatic lipid metabolism through protein kinase A-dependent sterol regulatory element binding protein1c degradation. Hepatology 60(3):844–857

[18] Liang Q, Xia W, Li W, Jiao J (2018) RNF20 controls astrocytic differentiation through epigenetic regulation of STAT3 in the developing brain . Cell Death Differ 25(2):294–306

[19] Liang X, Pan J, Cao C, Zhang L, Zhao Y, Fan Y, Li K, Tao C, Wang Y (2019) Transcriptional response of subcutaneous white adipose tissue to acute cold exposure in mice. Int J Mol Sci 20(16):3968

[20] Liu L,Jiang Q, Wang X, Zhang Y, Lin RC, Lam SM, Shui G, Zhou L, Li P, Wang Y(2014) Adipose-specific knockout of SEIPIN/ BSCL2 results in progressive lipodystrophy. Diabetes 63 (7):2320–2331

[21] Minsky N, Shema E, Field Y, Schuster M, Segal E, Oren M (2008) Monoubiquitinated H2B is associated with the transcribed region of highly expressed genes in human cells. Nat Cell Biol 10 (4):483–488

[22] Nakamura K, Kato A, Kobayashi J, Yanagihara H, Sakamoto S, Oliveira DV, Shimada M, Tauchi H, Suzuki H, Tashiro S (2011) Regulation of homologous recombination by RNF20- dependent H2B ubiquitination. Mol Cell 41(5):515–528

[23] Oelkrug R, Polymeropoulos ET, Jastroch M (2015) Brown adipose tissue: physiological function and evolutionary significance. J Comp Physiol B 185(6):587–606

[24] Pan D, Huang L, Zhu LJ, Zou T, Ou J, Zhou W, Wang YX (2015) Jmjd3-mediated H3K27me3 dynamics orchestrate brown fat development and regulate white fat plasticity. Dev Cell 35 (5):568–583

[25] Pan J,Tao C, Cao C, Zheng Q, Lam SM, Shui G, Liu X, Li K, Zhao J, Wang Y (2019) Adipose lipidomics and RNA-Seq analysis revealed the enhanced mitochondrial function in UCP1 knock-in pigs. Biochim Biophys Acta Mol Cell Biol Lipids 1864(10):1375–1383

[26] Phillips KJ (2019) Beige fat, adaptive thermogenesis, and its regulation by exercise and thyroid hormone. Biology (Basel) 8 (3):57

[27] Ren P, Sheng Z, Wang Y, Yi X, Zhou Q, Zhou J, Xiang S, Hu X, Zhang J (2014) RNF20 promotes the polyubiquitination and proteasome-dependent degradation of AP-2alpha protein. Acta Biochim Biophys Sin (Shanghai) 46(2):136–140

[28] Savage DB (2009) Mouse models of inherited lipodystrophy. Dis Model Mech 2(11–12):554–562

[29] Sethi G, Shanmugam MK, Arfuso F, Kumar AP (2018) Role of RNF20 in cancer development and progression- a comprehensive review. Biosci Rep 38(4):BSR20171287

[30] So CC, Ramachandran S, Martin A (2019) E3 ubiquitin ligases RNF20 and RNF40 are required for double-stranded break (DSB) repair: evidence for monoubiquitination of histone H2B lysine 120 as a novel axis of DSB signaling and repair. Mol Cell Biol 39 (8):488

[31] Tao C, Wang Y, Zhao Y, Pan J, Fan Y, Liang X,Cao C, Zhao J, Petris MJ, Li K, (2019) Adipocyte-specific disruption of ATPase copper transporting alpha in mice accelerates lipoatrophy. Diabetologia 62(12):2340–2353

[32] Tarcic O, Pateras IS, Cooks T,Shema E, Kanterman J, Ashkenazi H, Boocholez H, Hubert A, Rotkopf R, Baniyash M (2016) RNF20 links histone H2B ubiquitylation with inflammation and inflammation-associated cancer. Cell Rep 14(6):1462–1476

[33] Tseng YH, Cypess AM, Kahn CR (2010) Cellular bioenergetics as a target for obesity therapy. Nat Rev Drug Discov 9(6):465–482

[34] Van Marken Lichtenbelt W D, Vanhommerig JW, Smulders NM, Drossaerts JM, Kemerink GJ, Bouvy ND, Schrauwen P, Teule GJ (2009) Cold-activated brown adipose tissue in healthy men. N Engl J Med 360(15):1500–1508

[35] Vethantham V, Yang Y, Bowman C, Asp P,Lee JH, Skalnik DG, Dynlacht BD (2012) Dynamic loss of H2B ubiquitylation without corresponding changes in H3K4 trimethylation during myogenic differentiation. Mol Cell Biol 32(6):1044–1055

[36] Virtanen KA, Lidell ME, Orava J, Heglind M, Westergren R, Niemi T, Taittonen M, Laine J, Savisto NJ, Enerback S, (2009) Functional brown adipose tissue in healthy adults. N Engl J Med 360(15):1518–1525

[37] Vogel P,Read R, Hansen G, Wingert J, Dacosta CM, Buhring LM, Shadoan M (2011) Pathology of congenital generalized lipodystrophy in Agpat2−/− mice. Vet Pathol 48(3):642–654

[38] Woo Park J, Kim KB, Kim JY, Chae YC, Jeong OS, Seo SB (2015) RE-IIBP Methylates H3K79 and Induces MEIS1-mediated Apoptosis via H2BK120 Ubiquitination by RNF20. Sci Rep 5:12485

[39] Worden EJ, Wolberger C (2019) Activation and regulation of H2BUbiquitin-dependent histone methyltransferases. Curr Opin Struct Biol 59:98–106

[40] Wu C, Cui Y, Liu X, Zhang F,Lu LY, Yu X(2020) The RNF20/40 complex regulates p53-dependent gene transcription and mRNA splicing. J Mol Cell Biol 12(2):113–124

[41] Xu Z, Song Z, Li G, Tu H, Liu W, Liu Y, Wang P, Wang Y, Cui X, Liu C (2016) H2B ubiquitination regulates meiotic recombination by promoting chromatin relaxation. Nucleic Acids Res 44 (20):9681–9697

[42] Zingaretti MC, Crosta F, Vitali A, Guerrieri M, Frontini A, Cannon B, Nedergaard J, Cinti S (2009) The presence of UCP1 demonstrates that metabolically active adipose tissue in the neck of adult humans truly represents brown adipose tissue. FASEB J 23 (9):3113–3120

RIGHTS & PERMISSIONS

The Author(s)