Chiefari Eusebio, Arcidiacono Biagio, Possidente Katiuscia, Iiritano Stefania, Ventura Valeria, Pandolfo Rosantony, Brunetti Francesco Saverio, Greco Manfredi, Foti Daniela, Brunetti Antonio
Department of Health Sciences, University "Magna Græcia" of Catanzaro, Viale Europa (Loc. Germaneto), Catanzaro, Italy.
Department of Medical and Surgical Sciences, University "Magna Græcia" of Catanzaro, Viale Europa (Loc. Germaneto), Catanzaro, Italy.
PLoS One. 2013 Dec 18;8(12):e83969. doi: 10.1371/journal.pone.0083969. eCollection 2013.
The High-Mobility Group AT-Hook 1 (HMGA1) protein is an architectural transcription factor that binds to AT-rich sequences in the promoter region of DNA and functions as a specific cofactor for gene activation. Previously, we demonstrated that HMGA1 is a key regulator of the insulin receptor (INSR) gene and an important downstream target of the INSR signaling cascade. Moreover, from a pathogenic point of view, overexpression of HMGA1 has been associated with human cancer, whereas functional variants of the HMGA1 gene have been recently linked to type 2 diabetes mellitus and metabolic syndrome. However, despite of this biological and pathological relevance, the mechanisms that control HMGA1 gene expression remain unknown. In this study, to define the molecular mechanism(s) that regulate HMGA1 gene expression, the HMGA1 gene promoter was investigated by transient transfection of different cell lines, either before or after DNA and siRNA cotransfections. An octamer motif was identified as an important element of transcriptional regulation of this gene, the interaction of which with the octamer transcription factors Oct-1 and Oct-2 is crucial in modulating HMGA1 gene and protein expression. Additionally, we demonstrate that HMGA1 binds its own promoter and contributes to its transactivation by Oct-2 (but not Oct-1), supporting its role in an auto-regulatory circuit. Overall, our results provide insight into the transcriptional regulation of the HMGA1 gene, revealing a differential control exerted by both Oct-1 and Oct-2. Furthermore, they consistently support the hypothesis that a putative defect in Oct-1 and/or Oct-2, by affecting HMGA1 expression, may cause INSR dysfunction, leading to defects of the INSR signaling pathway.
高迁移率族AT钩蛋白1(HMGA1)是一种结构转录因子,可与DNA启动子区域富含AT的序列结合,并作为基因激活的特异性辅因子发挥作用。此前,我们证明HMGA1是胰岛素受体(INSR)基因的关键调节因子,也是INSR信号级联反应的重要下游靶点。此外,从致病角度来看,HMGA1的过表达与人类癌症有关,而HMGA1基因的功能变异最近与2型糖尿病和代谢综合征相关。然而,尽管具有这种生物学和病理学相关性,但控制HMGA1基因表达的机制仍然未知。在本研究中,为了确定调节HMGA1基因表达的分子机制,在DNA和siRNA共转染之前或之后,通过不同细胞系的瞬时转染对HMGA1基因启动子进行了研究。一个八聚体基序被确定为该基因转录调控的重要元件,其与八聚体转录因子Oct-1和Oct-2的相互作用对于调节HMGA1基因和蛋白表达至关重要。此外,我们证明HMGA1结合其自身启动子,并通过Oct-2(而非Oct-1)促进其反式激活,支持其在自调节回路中的作用。总体而言,我们的结果为HMGA1基因的转录调控提供了见解,揭示了Oct-1和Oct-2发挥的差异控制作用。此外,它们一致支持这样的假设,即Oct-1和/或Oct-2的假定缺陷通过影响HMGA1表达可能导致INSR功能障碍,进而导致INSR信号通路缺陷。