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17β-Estradiol-induced interaction of estrogen receptor α and human atrial essential myosin light chain modulates cardiac contractile function

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Abstract

Chronic increased workload of the human heart causes ventricular hypertrophy, re-expression of the atrial essential myosin light chain (hALC-1), and improved contractile function. Although hALC-1 is an important positive inotropic regulator of the human heart, little is known about its regulation. Therefore, we investigated the role of the sex hormone 17β-estradiol (E2) on hALC-1 gene expression, the underlying molecular mechanisms, and the impact of this regulatory process on cardiac contractile function. We showed that E2 attenuated hALC-1 expression in human atrial tissues of both sexes and in human ventricular AC16 cells. E2 induced the nuclear translocation of estrogen receptor alpha (ERα) and hALC-1 in AC16 cells, where they cooperatively regulate the transcriptional activity of hALC-1 gene promoter. E2-activated ERα required the estrogen response element (ERE) motif within the hALC-1 gene promoter to reduce its transcriptional activity (vehicle: 15.55 ± 4.80 vs. E2: 6.51 ± 3.69; ~2 fold). This inhibitory effect was potentiated in the presence of hALC-1 (vehicle: 11.13 ± 3.66 vs. E2: 2.18 ± 1.10; ~5 fold), and thus, hALC-1 acts as a co-repressor of ERα-mediated transcription. Yeast two-hybrid screening of a human heart cDNA library revealed that ERα interacts physically with hALC-1 in the presence of E2. This interaction was confirmed by Co-Immunoprecipitation and immunofluorescence in human atrium. As a further novel effect, we showed that chronic E2-treatment of adult mouse cardiomyocytes overexpressing hALC-1 resulted in reduced cell-shortening amplitude and twitching kinetics of these cells independent of Ca2+ activation levels. Together, our data showed that the expression of hALC-1 gene is, at least partly, regulated by E2/ERα, while hALC-1 acts as a co-repressor. The inotropic effect of hALC-1 overexpression in cardiomyocytes can be significantly repressed by E2.

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Abbreviations

aa:

Amino acid

Adv:

Adenovirus

ALC-1 (Myl4):

Atrial essential myosin light chain 4

β-Gal:

β-galactosidase

Co-IP:

Co-immunoprecipitation

DAPI:

4′,6-Diamidin-2-phenylindol

E2:

(17β-estradiol, estrogen)

EF:

Ejection fraction

ELC:

Essential myosin light chain

ERE:

Estrogen response element

ERα:

Estrogen receptor alpha

Eth:

Ethanol

FL:

Full length

IF:

Immunofluorescence

LUC:

Luciferase

MOI:

Multiplicity of infection

MyHC:

Myosin heavy chain

NPPA:

Atrial natriuretic peptide precursor A

VLC-1 (Myl3):

Ventricular essential myosin light chain

Y2H:

Yeast two-hybrid assay

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Acknowledgements

This work was supported by Friede Springer Herz-Stiftung [to SM] and Deutsche Forschungsgemeinschaft (DFG-FOR1054 to VRZ and SM). We thank Britta Fielitz, Arne Kuehne, Petra Domaing, and Karin Karczewski for excellent technical support. We also thank Prof. Pierre Chambon (INSERM, France) for the generous gift of ERα-pSG5 plasmid.

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    Authors and Affiliations

    1. Max-Delbrueck-Center for Molecular Medicine (MDC) in the Helmholtz Association, Robert-Roessle-Str. 10, 13125, Berlin, Germany

      Karolin Duft, Ahmed Abdelwahab, Cindy Schriever, Ingo Morano & Shokoufeh Mahmoodzadeh

    2. Institute of Gender in Medicine and Center for Cardiovascular Research, Charité Universitaetsmedizin Berlin, Berlin, Germany

      Miriam Schanz, Hang Pham, Georgios Kararigas, Elke Dworatzek & Vera Regitz-Zagrosek

    3. DZHK (German Center for Cardiovascular Research), partner site Berlin, Berlin, Germany

      Georgios Kararigas, Elke Dworatzek, Vera Regitz-Zagrosek, Ingo Morano & Shokoufeh Mahmoodzadeh

    4. Department of Radiation Oncology, Columbia University, New York, USA

      Mercy M. Davidson

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    1. Karolin Duft
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    Correspondence to Shokoufeh Mahmoodzadeh.

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    Ethical statements

    The use of human cardiac samples was approved by the Charité University Hospital review Ethics board (Berlin, Germany) and followed the rules of the Declaration of Helsinki. Samples were obtained with patient informed consent prior to the procedure.

    All procedures involving mice were approved by and conducted in accordance with the guidelines set out by the State Agency for Health and Social Affairs (LaGeSo, Berlin, Germany).

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    MM. Davidson is the holder of the patent on AC16 cells, and other authors have declared that no competing interests exist.

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    K. Duft and M. Schanz contributed equally to this work.

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    Duft, K., Schanz, M., Pham, H. et al. 17β-Estradiol-induced interaction of estrogen receptor α and human atrial essential myosin light chain modulates cardiac contractile function. Basic Res Cardiol 112, 1 (2017). https://doi.org/10.1007/s00395-016-0590-1

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