Abstract
Lysosome is a crucial organelle in charge of degrading proteins and damaged organelles to maintain cellular homeostasis. Transcription factor EB (TFEB) is the master transcription factor regulating lysosomal biogenesis and autophagy. Under external stimuli such as starvation, dephosphorylated TFEB transports into the nucleus to specifically recognize and bind to the coordinated lysosomal expression and regulation (CLEAR) elements at the promotors of autophagy and lysosomal biogenesis-related genes. The function of TFEB in the nucleus is fine regulated but the molecular mechanism is not fully elucidated. In this study, we discovered that miR-30b-5p, a small RNA which is known to regulate a series of genes through posttranscriptional regulation in the cytoplasm, was translocated into the nucleus, bound to the CLEAR elements, suppressed the transcription of TFEB-dependent downstream genes, and further inhibited the lysosomal biogenesis and the autophagic flux; meanwhile, knocking out the endogenous miR-30b-5p by CRISPR/Cas9 technique significantly increased the TFEB-mediated transactivation, resulting in the increased expression of autophagy and lysosomal biogenesis-related genes. Overexpressing miR-30b-5p in mice livers showed a decrease in lysosomal biogenesis and autophagy. These in vitro and in vivo data indicate that miR-30b-5p may inhibit the TFEB-dependent transactivation by binding to the CLEAR elements in the nucleus to regulate the lysosomal biogenesis and autophagy. This novel mechanism of nuclear miRNA regulating gene transcription is conducive to further elucidating the roles of miRNAs in the lysosomal physiological functions and helps to understand the pathogenesis of abnormal autophagy-related diseases.
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Acknowledgements
We thank Yue Yin and Ping Wu (National Center for Protein Science—Shanghai) for mass spectrometry, Yizheng Wang (The Brain Science Center, Beijing Institute of Basic Medical Sciences) and Kai Fu (Xiangya Hospital Central South University) for helpful comments. This work was supported by the National Natural Science Foundation of China (81701261 and 81771890), the National Science & Technology Major Project “Key New Drug Creation and Manufacturing Program,” China (2018ZX09101001-003-007, 2018ZX09201017-004, 2019ZX09732002-013, and 2018ZX09711002-010-001), the Open Research Fund of State Key Laboratory of Transient Optics and Photonics (SKLST201806), the Fundamental and Open Research Funds from the State Key Laboratory of Drug Research (SIMM1903ZZ-05 and SIMM2004KF-02), Shanghai Committee of Science and Technology of China under Grant No.18DZ2290200.
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Guo, H., Pu, M., Tai, Y. et al. Nuclear miR-30b-5p suppresses TFEB-mediated lysosomal biogenesis and autophagy. Cell Death Differ 28, 320–336 (2021). https://doi.org/10.1038/s41418-020-0602-4
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DOI: https://doi.org/10.1038/s41418-020-0602-4
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