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mRNA m5C inhibits adipogenesis and promotes myogenesis by respectively facilitating YBX2 and SMO mRNA export in ALYREF-m5C manner

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Abstract

Although 5-methylcytosine (m5C) has been identified as a novel and abundant mRNA modification and associated with energy metabolism, its regulation function in adipose tissue and skeletal muscle is still limited. This study aimed at investigating the effect of mRNA m5C on adipogenesis and myogenesis using Jinhua pigs (J), Yorkshire pigs (Y) and their hybrids Yorkshire–Jinhua pigs (YJ). We found that Y grow faster than J and YJ, while fatness-related characteristics observed in Y were lower than those of J and YJ. Besides, total mRNA m5C levels and expression rates of NSUN2 were higher both in backfat layer (BL) and longissimus dorsi muscle (LDM) of Y compared to J and YJ, suggesting that higher mRNA m5C levels positively correlate with lower fat and higher muscle mass. RNA bisulfite sequencing profiling of m5C revealed tissue-specific and dynamic features in pigs. Functionally, hyper-methylated m5C-containing genes were enriched in pathways linked to impaired adipogenesis and enhanced myogenesis. In in vitro, m5C inhibited lipid accumulation and promoted myogenic differentiation. Furthermore, YBX2 and SMO were identified as m5C targets. Mechanistically, YBX2 and SMO mRNAs with m5C modification were recognized and exported into the cytoplasm from the nucleus by ALYREF, thus leading to increased YBX2 and SMO protein expression and thereby inhibiting adipogenesis and promoting myogenesis, respectively. Our work uncovered the critical role of mRNA m5C in regulating adipogenesis and myogenesis via ALYREF-m5C-YBX2 and ALYREF-m5C-SMO manners, providing a potential therapeutic target in the prevention and treatment of obesity, skeletal muscle dysfunction and metabolic disorder diseases.

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Availability of data and material

All data generated or analyzed during this study are included in this article. The datasets used during the current study have been deposited in the Genome Sequence Archive under the accession number CRA005591 linked to the project PRJCA007533.

Code availability

The authors declare that the codes for data analyses from this study are available from the corresponding authors on reasonable request.

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Acknowledgements

We would like to thank the reviewers for their constructive comments.

Funding

This work is supported by the Natural Science Foundation of Zhejiang Province (LZ22C170002); the National Natural Science Foundation of China (U21A20249); the Fundamental Research Funds for the Central Universities (2019XZZX003-13); the National Key R & D Program (2018YFD0500400); the National Natural Science Foundation of China (31572413); the Natural Science Foundation of Zhejiang Province (LZ17C170001).

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Authors and Affiliations

  1. College of Animal Sciences, Zhejiang University, Hangzhou, Zhejiang, China

    Youhua Liu, Ruifan Wu, Xing Liao, Botao Zeng, Chaoqun Huang, Yaojun Luo, Yuxi Liu, Yushi Chen, Wei Chen, Jiaqi Liu, Qin Jiang, Yuanling Zhao, Zhen Bi, Guanqun Guo, Yongxi Yao, Teresa G. Valencak, Yizhen Wang & Xinxia Wang

  2. Key Laboratory of Animal Nutrition and Feed Sciences, Ministry of Agriculture, Hangzhou, China

    Youhua Liu, Ruifan Wu, Xing Liao, Botao Zeng, Chaoqun Huang, Yaojun Luo, Yuxi Liu, Yushi Chen, Wei Chen, Jiaqi Liu, Qin Jiang, Yuanling Zhao, Zhen Bi, Guanqun Guo, Yongxi Yao, Yizhen Wang & Xinxia Wang

  3. Laboratory of Molecular Animal Nutrition (Zhejiang University), Ministry of Education, Hangzhou, China

    Youhua Liu, Ruifan Wu, Xing Liao, Botao Zeng, Chaoqun Huang, Yaojun Luo, Yuxi Liu, Yushi Chen, Wei Chen, Jiaqi Liu, Qin Jiang, Yuanling Zhao, Zhen Bi, Guanqun Guo, Yongxi Yao, Yizhen Wang & Xinxia Wang

  4. Key Laboratory of Animal Feed and Nutrition of Zhejiang Province, Hangzhou, Zhejiang, China

    Youhua Liu, Ruifan Wu, Xing Liao, Botao Zeng, Chaoqun Huang, Yaojun Luo, Yuxi Liu, Yushi Chen, Wei Chen, Jiaqi Liu, Qin Jiang, Yuanling Zhao, Zhen Bi, Guanqun Guo, Yongxi Yao, Yizhen Wang & Xinxia Wang

  5. CAS Key Laboratory of Genomic and Precision Medicine, Collaborative Innovation Center of Genetics and Development, College of Future Technology, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China

    Ying Yang, Chun-chun Gao & Xiao Han

  6. China National Center for Bioinformation, Hangzhou, China

    Ying Yang, Chun-chun Gao & Xiao Han

  7. Institute of Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, China

    Ying Yang, Chun-chun Gao & Xiao Han

  8. Jinhua Academy of Agricultural Sciences, Jinhua, China

    Yun Xiang & Xiaojun Zhang

Authors
  1. Youhua Liu
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  2. Ying Yang
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  3. Ruifan Wu
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  21. Teresa G. Valencak
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  22. Yizhen Wang
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  23. Xinxia Wang
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Contributions

YHL, YY, RFW, XL, BTZ, CQH, YJL, YXL, YSC, WC, QJL, QJ, YLZ, ZB, GQG and YXY performed experiments under the supervision of XXW. YHL, CCG and XH performed bioinformatics analysis of sequence data. YHL wrote the manuscript under the supervision of XXW. YX, XJZ, YZW and XXW designed the project and XXW provided the final approval of the manuscript. Teresa G. Valencak reorganized the writing and figure layout, and revised the typo errors and grammar mistakes throughout our manuscript.

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Correspondence to Xinxia Wang.

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Sample collection in this study was performed according to the guidelines for the care and use of experimental animals established by the Ministry of Agriculture of China. All procedures were approved by the Committee on Animal Care and Use and Committee on the Ethics of Animal Experiments of Zhejiang University (ZJU2015-458-09).

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Liu, Y., Yang, Y., Wu, R. et al. mRNA m5C inhibits adipogenesis and promotes myogenesis by respectively facilitating YBX2 and SMO mRNA export in ALYREF-m5C manner. Cell. Mol. Life Sci. 79, 481 (2022). https://doi.org/10.1007/s00018-022-04474-0

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