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Novel Link of Anti-apoptotic ATF3 with Pro-apoptotic CTMP in the Ischemic Brain

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Abstract

Activating transcription factor 3 (ATF3) is a stress-induced transcription factor with diverse functions under disease states in multiple cell types. ATF3 has neuroprotective action against cerebral ischemia, which may involve caspase 3. However, the molecular mechanisms underlying ATF3 regulation of apoptosis are largely unknown. Here, we used gain- and loss-of-function and rescue approaches to demonstrate ATF3 attenuating hypoxic neuronal apoptosis. As well, the protective effect of ATF3 was mediated by downregulation of carboxyl-terminal modulator protein (CTMP), a pro-apoptotic factor that inhibits the anti-apoptotic Akt/PKB cascade. ATF3 (1) downregulated the mRNA and protein levels of CTMP; (2) its temporal expression pattern was reciprocal to that of CTMP; and (3) nuclear localization suggested that ATF3 may regulate CTMP transcription following hypoxic insult. Reporter assays demonstrated that ATF3 suppressed CTMP transcription, whereas ATF3 fusion with VP16, converting ATF3 to transcriptional activator, boosted CTMP transcription. By contrast, NF-κB increased CTMP transcription, and degradation-resistant IκBα decreased CTMP transcription. ChIP assays further confirmed that binding of ATF3 to the ATF/CREB site hindered NF-κB binding to the CTMP promoter, which repressed CTMP expression. Furthermore, CTMP siRNA treatment reduced hypoxic neuronal apoptosis by increasing p-Akt (Ser473) levels and leaving the upstream ATF3 level unchanged. We have identified an endogenous neuroprotective ATF3→CTMP signal cascade that may be a therapeutic target for reducing ischemic brain injury.

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Acknowledgments

We thank Yu-Chang Chen and Yen-Ling Chen for the technical support. We thank the Institute of Biomedical Sciences Instrumental Core Facility, Academia Sinica for confocal microscopy assistance. This work was supported by grants from the National Science Council and Academia Sinica, Taiwan. The authors had full access to and take full responsibility for the integrity of the data. All authors have read and agree to the manuscript as written.

Conflict of Interests

The authors declare no conflict of interest.

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Authors and Affiliations

  1. Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan

    Chien-Yu Huang, Yuan-Soon Ho & Teng-Nan Lin

  2. Institute of Biomedical Sciences, Academia Sinica, Rm 404, IBMS, Academia Sinica, 11529, Taipei, Taiwan, Republic of China

    Chien-Yu Huang, Jin-Jer Chen, Jui-Sheng Wu, Hsin-Da Tsai, Yu-Ting Yan & Teng-Nan Lin

  3. Department of Internal Medicine, Division of Cardiology, China Medical University Hospital, Taichung, Taiwan

    Jin-Jer Chen

  4. Graduate Institute of Clinical Medical Science, China Medical University, Taichung, Taiwan

    Jin-Jer Chen & Chung Y. Hsu

  5. Department of Physiology, College of Medicine, Taipei Medical University, Taichung, Taiwan

    Heng Lin

  6. Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan

    Teng-Nan Lin

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  1. Chien-Yu Huang
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Huang, CY., Chen, JJ., Wu, JS. et al. Novel Link of Anti-apoptotic ATF3 with Pro-apoptotic CTMP in the Ischemic Brain. Mol Neurobiol 51, 543–557 (2015). https://doi.org/10.1007/s12035-014-8710-0

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