de Francisco Patricia, Amaro Francisco, Martín-González Ana, Gutiérrez Juan Carlos
Departamento de Genética, Fisiología y Microbiología, Facultad de Ciencias Biológicas, Universidad Complutense de Madrid, Madrid, Spain.
Front Genet. 2018 Oct 23;9:459. doi: 10.3389/fgene.2018.00459. eCollection 2018.
Metallothioneins (MT) are multi-stress proteins mainly involved in metal detoxification. MT gene expression is normally induced by a broad variety of stimulus and its gene expression regulation mainly occurs at a transcriptional level. Conserved motifs in the MT promoters have been described. These motifs show a consensus sequence very similar to AP-1 sites, and bZIP type transcription factors might participate in the MT gene expression regulation. In this research work, we characterize four AP-1 transcription factors in each of four different analyzed species, detecting a high conservation among them. Each AP-1 molecule has its counterpart in the other three species. A comparative qRT-PCR analysis of these AP-1 genes have been carried out in different strains (including metal-adapted, knockout and/or knockdown strains among others), and under different metal-stress conditions (1 or 24 h Cd, Cu, or Pb treatments). The possible interaction of these transcription factors with the conserved AP-1 motifs present in MT promoters has been corroborated by protein-DNA interaction experiments. Certain connection between the expression patterns of the bZIP and MT genes seems to exist. For the first time, and based on our findings, a possible gene expression regulation model including both AP-1 transcription factors and MT genes from the ciliate has been elaborated.
金属硫蛋白(MT)是主要参与金属解毒的多应激蛋白。MT基因表达通常由多种刺激诱导,其基因表达调控主要发生在转录水平。MT启动子中的保守基序已被描述。这些基序显示出与AP-1位点非常相似的共有序列,并且bZIP型转录因子可能参与MT基因表达调控。在这项研究工作中,我们对四个不同分析物种中的每一个中的四个AP-1转录因子进行了表征,检测到它们之间具有高度保守性。每个AP-1分子在其他三个物种中都有其对应物。已经在不同菌株(包括金属适应株、基因敲除和/或基因敲低株等)以及不同金属应激条件(1或24小时镉、铜或铅处理)下对这些AP-1基因进行了比较定量逆转录聚合酶链反应(qRT-PCR)分析。蛋白质-DNA相互作用实验证实了这些转录因子与MT启动子中存在的保守AP-1基序之间可能的相互作用。bZIP和MT基因的表达模式之间似乎存在某种联系。基于我们的发现,首次阐述了一个可能的基因表达调控模型,该模型包括来自纤毛虫的AP-1转录因子和MT基因。