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JNK-dependent phosphorylation and nuclear translocation of EGR-1 promotes cardiomyocyte apoptosis

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Abstract

Myocardial apoptosis induced by myocardial ischemia and hyperlipemia are the main causes of high mortality of cardiovascular diseases. It is not clear whether there is a common mechanism responsible for these two kinds of cardiomyocyte apoptosis. Previous studies demonstrated that early growth response protein 1 (EGR-1) has a pro-apoptotic effect on cardiomyocytes under various stress conditions. Here, we found that EGR-1 is also involved in cardiomyocyte apoptosis induced by both ischemia and high-fat, but how EGR-1 enters the nucleus and whether nuclear EGR-1 (nEGR-1) has a universal effect on cardiomyocyte apoptosis are still unknown. By analyzing the phosphorylation sites and nucleation information of EGR-1, we constructed different mutant plasmids to confirm that the nucleus location of EGR-1 requires Ser501 phosphorylation and regulated by JNK. Furthermore, the pro-apoptotic effect of nEGR-1 was further explored through genetic methods. The results showed that EGR-1 positively regulates the mRNA levels of apoptosis-related proteins (ATF2, CTCF, HAND2, ELK1), which may be the downstream targets of EGR-1 to promote the cardiomyocyte apoptosis. Our research announced the universal pro-apoptotic function of nEGR-1 and explored the mechanism of its nucleus location in cardiomyocytes, providing a new target for the “homotherapy for heteropathy” to cardiovascular diseases.

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Abbreviations

ATF2:

Cyclic AMP-dependent transcription factor 2

CTCF:

Transcriptional repressor

EGR-1:

Early growth response protein 1

nEGR-1:

Nuclear EGR-1

ELK1:

ETS domain-containing protein 1

HAND2:

Heart- and neural crest derivatives-expressed protein 2

JNK:

Mitogen-activated protein kinase 8

MAPK:

Mitogen-activated protein kinase

NLS:

Nuclear localization signals

PA:

Palmitic acid

SD:

Serum deprivation

SPC:

Sphingosylphosphorylcholine

TF:

Transcription factor

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Acknowledgements

This work was supported by grants of the National Natural Science Foundation of China (Grant No. 31671180). We thank Sen Wang, Haiyan Yu, Xiaomin Zhao, YuYu Guo from State Key Laboratory of Microbial Technology, Shandong University for the assistance in microimaging of Fluorescence microscope analysis.

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Authors and Affiliations

  1. Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Science, Shandong University, Qingdao, 266237, People’s Republic of China

    Jinrun Zhou, Yujuan Yao, Jiaojiao Zhang, Zhaohui Wang, Tianshu Zheng, Yao Lu, Weihua Kong & Jing Zhao

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  1. Jinrun Zhou
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  2. Yujuan Yao
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  3. Jiaojiao Zhang
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  8. Jing Zhao
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Contributions

The study was designed by Jing Zhao. Jinrun Zhou and Yujuan Yao carried out the experiments, Jiaojiao Zhang, Zhaohui Wang, Tianshu Zheng and Yao Lu analyzed data. Jing Zhao and Jinrun Zhou wrote the paper. Jing Zhan and Weihua Kong checked and finalized the manuscript. All authors read and approved the final manuscript.

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Correspondence to Jing Zhao.

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10495_2022_1714_MOESM1_ESM.tif

Supplementary file1 (TIF 1666 KB) Fig. S1 A, B H9c2 cells were serum deprived and treated with SPC (5 μM) for 12 h, the levels of apoptosis were detected by TUNEL analysis. C, D H9c2 cells were treated with BSA and PA (0.4 mM) for 12 h, the levels of apoptosis were detected by TUNEL analysis. Data are expressed as the mean ± S.D. *p < 0.05 and **p < 0.01; n = 3

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Zhou, J., Yao, Y., Zhang, J. et al. JNK-dependent phosphorylation and nuclear translocation of EGR-1 promotes cardiomyocyte apoptosis. Apoptosis 27, 246–260 (2022). https://doi.org/10.1007/s10495-022-01714-3

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