Cu^(2+)胁迫后青海湖裸鲤DNAJC2及其相关基因的表达分析

Expression Analysis of DNAJC2 and Related Genes in Gymnocypris przewalskii Under Cu^(2+)Stress

为进一步明确青海湖裸鲤(Gymnocypris przewalskii)应对胁迫反应的分子机制,并挖掘有重要功能的基因,本研究克隆了青海湖裸鲤中一个编码J结构域的基因DNAJC2,并分析了其相关基因对Cu^(2+)胁迫的应答反应。研究发现,青海湖裸鲤DNAJC2基因的开放阅读框为1866 bp,共编码621个氨基酸,5′-UTR非翻译区为108 bp,3′-UTR非翻译区为130 bp。DNAJC2在青海湖裸鲤肝脏、脑和鳃组织中高表达。与空白对照组相比,随着Cu^(2+)浓度和胁迫时间的增加,在鳃和肾脏组织中,Grp78、ID1和DNAJC2基因以表达上调的方式参与应激反应,而Hspa14、Hyou1、RARα和XPC基因以表达下调或者上调后又恢复至正常水平的方式参与应激反应;在肝脏中,Hspa14和ID1基因以表达上调的方式参与应激反应,而Grp78、DNAJC2、CDK8和XPC基因以表达上调或下调后又下调或上调的方式参与应激反应,Hyou1和RARα基因在低浓度下以表达上调的方式参与应激反应,但在高浓度下以表达下调后又上调的方式参与应激反应。以上结果表明,Hspa14、Grp78、ID1和DNAJC2基因参与了机体应激外源性胁迫的生物过程并发挥了重金属解毒作用,且外源性Cu^(2+)胁迫可影响Hyou1、CDK8、RARα和XPC调控的靶基因转录和蛋白质的合成。

To further clarify the molecular mechanism of Gymnocypris przewalskii in response to stress and explore gene functions,this study cloned DNAJC2,a gene encoding J domain,and analyzed the response patterns of its related genes to Cu^(2+)stress.The study found that the open reading frame of DNAJC2 was 1866 bp,encoding 621 amino acids,the untranslated region of the 5′-UTR was 108 bp,and the untranslated region of the 3′-UTR was 130 bp.DNAJC2 was higher expressed in the livers,brains,and gills of G.przewalskii.Compared with the blank control group,with the increase of Cu^(2+)concentration and the prolongation of stress time,gene Grp78,ID1,and DNAJC2 were above regulated in gills and kidneys,while gene Hspa14,Hyou1,RARα,and XPC were down-regulated or up-regulated and then returned to normal levels.In the liver,gene Hspa14 and ID1 were above regulated to participate in the stress response,while gene Grp78,DNAJC2,CDK8,and XPC were up-regulated or down-regulated and then down-regulated or up-regulated.Gene Hyou1 and RARαparticipated in stress response in an up-regulated way at low concentration,but down-regulated and then up-regulated at high concentration.These results indicated that gene Hspa14,Grp78,ID1,and DNAJC2 were involved in the biological process of exogenous stress and the cellular detoxification of heavy metals,and exogenous Cu^(2+)stress could affect the transcription of target genes and protein synthesis regulated by Hyou1,CDK8,RARα,and XPC.