RNF20及其介导的组蛋白H2B单泛素化对小鼠棕色脂肪细胞分化的影响

doi:10.11843/j.issn.0366-6964.2020.04.008

Abstract

Abstract: The objective of this study was to investigate the effect of RNF20 and it-mediated monoubiquitination of histone H2B at lysine120 (H2Bub) on brown adipocytes differentiation in mice. In this study, the expression of RNF20 and it-mediated H2Bub level in brown adipose tissue (BAT) from 1-day-old and 2-month-old male C57BL/6 mice were measured by Western blot (n=3). The brown preadipocytes were primarily isolated by collagenase digestion from 1-day-old mice. The brown preadipocytes and C3H10T1/2 cell lines were differentiated into mature adipocytes, respectively. Oil Red O staining was used to detect the adipogenic differentiation efficiency. The expression of RNF20 and it-mediated H2Bub level were detected in non-differentiated and differentiated cells (0 and 8 d) by Western blot. Furthermore, the siRNAs were applied to knockdown the expression of Rnf20 gene in C3H10T1/2 cells and the Oil Red O staining was used to detect adipogenic differentiation efficiency. qPCR and Western blot were used to detect the efficiency of interfering Rnf20 gene and it-mediated H2Bub level. The results showed that the expression of RNF20 and it-mediated H2Bub level were significantly higher in BAT from 2-month-old mice than those from 1-day-old mice. The expression of adipocyte differentiation marker proteins, PPARγ and CEBPα, and RNF20 and H2Bub level significantly increased in differentiated brown adipocytes and C3H10T1/2 cells. In addition, knockdown of Rnf20 gene in C3H10T1/2 cells significantly decreased the expression of RNF20 and it-mediated H2Bub level and reduced lipid droplets deposition in differentiated cells compared to those from negative control group. In summary, RNF20 was essential for mouse brown adipocyte differentiation. Knockdown of Rnf20 gene resulted in the decrease of H2Bub level and decreased the adipogenic differentiation efficiency of C3H10T1/2 cells. The study result adds more data to the epigenetic regulation of mouse brown adipocytes differentiation and provides new genetic material for further understanding the differentiation of animal brown adipocytes.

Key words: Rnf20 gene, H2Bub, brown adipocytes, adipogenic differentiation, mice

CLC Number:

Q953

LIANG Xiaojuan, TAO Cong, ZHAO Ying, WANG Chao, LIU Lulu, WANG Yanfang. Effect of RNF20 and It-mediated Monoubiquitination of Histone H2B at Lysine 120 (H2Bub) on Brown Adipocyte Differentiation in Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(4): 722-731.

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Effect of RNF20 and It-mediated Monoubiquitination of Histone H2B at Lysine 120 (H2Bub) on Brown Adipocyte Differentiation in Mice

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