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Down-regulation of SM22/transgelin gene expression during H9c2 cells differentiation

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Abstract

The embryonic rat ventricle H9c2 cells maintain a proliferative state (P condition) in the presence of 10% FCS. However, by reducing serum concentration and in the presence of retinol acetate, proliferation is stopped, myogenic transdifferentiation is inhibited while cardiac differentiation is preserved (D condition). Two-dimensional gel electrophoresis and mass spectrometry analysis was used to define the modifications of the nuclear proteome occurring during the P-to-D transition. Among the proteins observed as modified, a reduced expression of the SM22/transgelin protein was associated with the D state. Also SM22 mRNA levels were reduced during P-to-D transition. Cell transfection experiments indicated that this decrease was partially due to a reduction of the SM22 promoter activity. GATA-4 had a repressive effect on SM22 promoter activity. Thus, since GATA-4 is known as a target of retinoids and may act as a transcriptional repressor, a mechanism to explain the SM22 reduction during the P-to-D transition is tentatively proposed. Immunohistochemical studies on heart cells confirmed the nuclear localization of SM22. Moreover, a differential expression of this protein in different districts of the human heart embryo was detected. Therefore, these data suggest that SM22 expression is regulated during heart development.

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Abbreviations

2-DE:

Bi-dimensional electrophoresis

MALDI-TOF:

Matrix assisted laser desorption ionization-time of flight

MS:

Mass spectrometry

μLC-ESI-IT-MS/MS:

Micro-liquid chromatography-electrospray-ion trap-tandem mass spectrometry

SM22:

Transgelin

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Acknowledgments

This work is funded by grants from MIUR to GD (PRIN No. 2007N8P32H_002), GT (FIRB No. RBRN07BMCT—Italian Human ProteomeNet) and AS (MERIT) and from CNR (AG.P04.015 and RSTL 862). Expression vector GATA-4 was provided by Prof. D. Salvatore (University Federico II, Naples).

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Authors and Affiliations

  1. Dipartimento di Scienze e Tecnologie Biomediche, Università di Udine, Piazzale Kolbe 1, 33100, Udine, Italy

    Elisa Bregant, Renata Lonigro, Nadia Passon, Gianluca Tell & Giuseppe Damante

  2. Proteomics and Mass Spectrometry Laboratory, ISPAAM, National Research Council, Naples, Italy

    Giovanni Renzone & Andrea Scaloni

  3. Dipartimento di Ricerche Mediche e Morfologiche, Università di Udine, Udine, Italy

    Carla Di Loreto & Maura Pandolfi

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  1. Elisa Bregant
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  2. Giovanni Renzone
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Correspondence to Giuseppe Damante.

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Bregant, E., Renzone, G., Lonigro, R. et al. Down-regulation of SM22/transgelin gene expression during H9c2 cells differentiation. Mol Cell Biochem 327, 145–152 (2009). https://doi.org/10.1007/s11010-009-0052-2

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  • DOI: https://doi.org/10.1007/s11010-009-0052-2

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