Identification of a potential tissue-specific biomarker cathepsin L-like gene from the planarian Dugesia japonica: Molecular cloning, characterization, and expression in response to heavy metal exposure
Identification of a potential tissue-specific biomarker cathepsin L-like gene from the planarian Dugesia japonica: Molecular cloning, characterization, and expression in response to heavy metal exposure
复制标题
从涡虫 Dugesia japonica 中鉴定潜在的组织特异性生物标志物组织蛋白酶 L 样基因：重金属暴露响应的分子克隆、表征和表达
DOI：
10.1016/j.ecoenv.2019.04.079
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发表时间：
2019
影响因子：
6.8
通讯作者：
De-Zeng Liu
中科院分区：
文献类型：
--
作者：
Ke-Xue Ma;Ge-Ge Song;Meng Wu;He-Cai Zhang;Guang-Wen Chen;De-Zeng Liu
重金属污染是一个全球性的健康问题，影响世界各地的人们，探索敏感的生物标志物来评估重金属的毒性是研究人员的重要工作。组织蛋白酶L作为一种组织特异性生物标志物，可用于评价环境污染物的生物学效应，但尚未受到重视。本研究利用cDNA末端快速扩增（RACE）技术克隆了日本三角涡虫（Dugesia japonica）组织蛋白酶L基因的全长cDNA（命名为DjCatL）。DjCatL的cDNA序列为1161 bp，编码346个氨基酸，分子量为39.03 kDa。序列分析表明，DjCatL含有高度保守的ERF/WNIN、GNFD和GCXGG基序，这些基序是组织蛋白酶L蛋白家族的特征。全膜原位杂交（whole-mountin situhybridization，WISH）结果表明，DjCatL基因的转录本特异性地分布在肠道系统中，提示该基因与真涡虫的食物消化有关。定量聚合酶链反应（qPCR）和WISH结果均表明，重金属（Cd 2+、Hg 2+和Cu 2+）对DjCatL基因转录水平有明显的抑制作用，且呈剂量依赖性。因此，组织蛋白酶L可以作为一种组织特异性生物标志物，用于评价水环境中的重金属污染。
Heavy metal pollution is a global health issue affecting people worldwide, and the exploration of sensitive biomarkers to assess the toxicity of heavy metals is an important work for researchers. Cathepsin L, role as a tissue-specific biomarker to assess the biological effects of environmental pollutants, has not received much attention. In this work, the full-length cDNA of cathepsin L gene from the planarian Dugesia japonica (designatedDjCatL) was cloned by rapid amplification of cDNA ends (RACE) technique. The cDNA sequence ofDjCatLis 1161 bp, which encodes a protein of 346 amino acids with a molecular weight of 39.03 kDa. Sequence analysis revealed that DjCatL contains highly conserved ERF/WNIN, GNFD, and GCXGG motifs, which are the features of the cathepsin L protein family. Whole-mountin situhybridization (WISH) results revealed that the transcripts ofDjCatLare specifically distributed in the intestinal system, suggesting that this gene is related to food digestion in planarians. Both quantitative polymerase chain reaction (qPCR) and WISH results revealed that the transcriptional levels ofDjCatLare inhibited significantly by heavy metal (Cd2+, Hg2+, and Cu2+) exposure in a dose-dependent manner. Therefore, we proposed that cathepsin L can be used as a tissue-specific biomarker to assess the heavy metal pollution in the aquatic environment.