School of Biosciences, Cardiff University, Cardiff CF10 3AX, Wales, UK.
Ecotoxicology. 2012 Jan;21(1):124-38. doi: 10.1007/s10646-011-0772-4. Epub 2011 Sep 11.
The macro-alga Fucus vesiculosus has a broad global and estuarine distribution and exhibits exceptional resistance to toxic metals, the molecular basis of which is poorly understood. To address this issue a cDNA library was constructed from an environmental isolate of F. vesiculosus growing in an area with chronic copper pollution. Characterisation of this library led to the identification of a cDNA encoding a protein known to be synthesised in response to toxicity, a full length 14-3-3 exhibiting a 71% identity to human/mouse epsilon isoform, 70-71% identity to yeast BMH1/2 and 95 and 71% identity to the Ectocarpus siliculosus 14-3-3 isoforms 1 and 2 respectively. Preliminary characterisation of the expression profile of the 14-3-3 indicated concentration- and time-dependent inductions on acute exposure of F. vesiculosus of copper (3-30 μg/l). Higher concentrations of copper (≥150 μg/l) did not elicit significant induction of the 14-3-3 gene compared with the control even though levels of both intracellular copper and the expression of a cytosolic metal chaperone, metallothionein, continued to rise. Analysis of gene expression within environmental isolates demonstrated up-regulation of the 14-3-3 gene associated with the known copper pollution gradient. Here we report for the first time, identification of a gene encoding a putative 14-3-3 protein in a multicellular alga and provide preliminary evidence to link the induction of this 14-3-3 gene to copper exposure in this alga. Interestingly, the threshold exposure profile may be associated with a decrease in the organism's ability to control copper influx so that it perceives copper as a toxic response.
大型褐藻泡叶藻在全球和河口广泛分布,并表现出对有毒金属的特殊抗性,但对其分子基础知之甚少。为了解决这个问题,从生长在慢性铜污染地区的环境分离物中构建了泡叶藻 cDNA 文库。该文库的特征分析导致鉴定出一个编码已知对毒性有反应的蛋白质的 cDNA,一个全长 14-3-3,与人/鼠 ε 同工型具有 71%的同一性,与酵母 BMH1/2 具有 70-71%的同一性,与 Ectocarpus siliculosus 14-3-3 同工型 1 和 2 分别具有 95%和 71%的同一性。14-3-3 的表达谱的初步特征表明,铜(3-30μg/l)急性暴露下泡叶藻的浓度和时间依赖性诱导。与对照相比,即使细胞内铜水平和胞质金属伴侣金属硫蛋白的表达继续上升,较高浓度的铜(≥150μg/l)并没有显著诱导 14-3-3 基因。对环境分离物中的基因表达分析表明,与已知的铜污染梯度相关,14-3-3 基因上调。在这里,我们首次报道了在多细胞藻类中鉴定出编码假定的 14-3-3 蛋白的基因,并提供了初步证据表明,该 14-3-3 基因的诱导与该藻类中铜暴露有关。有趣的是,阈值暴露谱可能与生物体控制铜流入的能力下降有关,因此它将铜视为一种毒性反应。