Bregant Elisa, Renzone Giovanni, Lonigro Renata, Passon Nadia, Di Loreto Carla, Pandolfi Maura, Scaloni Andrea, Tell Gianluca, Damante Giuseppe
Dipartimento di Scienze e Tecnologie Biomediche, Università di Udine, Piazzale Kolbe 1, 33100 Udine, Italy.
Mol Cell Biochem. 2009 Jul;327(1-2):145-52. doi: 10.1007/s11010-009-0052-2. Epub 2009 Feb 18.
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.
胚胎大鼠心室H9c2细胞在10%胎牛血清(FCS)存在的情况下维持增殖状态(P状态)。然而,通过降低血清浓度并在醋酸视黄醇存在的情况下,细胞增殖停止,成肌转分化受到抑制,而心脏分化得以保留(D状态)。二维凝胶电泳和质谱分析被用于确定从P状态到D状态转变过程中发生的核蛋白质组修饰。在观察到发生修饰的蛋白质中,SM22/转胶蛋白的表达降低与D状态相关。在从P状态到D状态的转变过程中,SM22 mRNA水平也降低。细胞转染实验表明,这种降低部分归因于SM22启动子活性的降低。GATA-4对SM22启动子活性具有抑制作用。因此,由于GATA-4是已知的类维生素A靶点且可能作为转录抑制因子,初步提出了一种解释从P状态到D状态转变过程中SM22降低的机制。对心脏细胞的免疫组织化学研究证实了SM22的核定位。此外,还检测到该蛋白在人类心脏胚胎不同区域的差异表达。因此,这些数据表明SM22的表达在心脏发育过程中受到调控。