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纳米氧化铈暴露下大型溞MAPK基因家族的表达分析
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Abstract:
MAPK基因家族广泛参与生物体细胞内的各种生命过程，与生物对环境胁迫的应答相关。然而，在纳米氧化铈(Nano cerium dioxide, nCeO2)暴露下大型溞MAPK基因家族成员的作用机制还不清楚。本文利用BLAST、HMM和SMART等方法在大型溞基因组中鉴定MAPK基因家族成员，分析大型溞各个MAPK基因的理化性质、染色体定位、结构域、保守基序、系统发生关系等，并利用qPCR分析不同浓度、不同时间nCeO2暴露下各个MAPK基因家族成员的表达模式。结果鉴定出14个大型溞的MAPK基因，分布于8条染色体上。MAPK基因家族编码的蛋白质序列长度为361~1546 aa，分子量为38.93~77.02 kDa，等电点在5.3~9.51。12个MAPK蛋白具有S_TKc结构域，4个保守基序。系统发生分析显示，大型溞的14个MAPK基因分别属于4个亚家族，基因序列在进化过程中具有较高的保守性。在50和150 mg/L nCeO2暴露48 h后，大型溞的7个MAPK基因DmMAPK-p38b、DmMAPK4、DmMAP2K4、DmMAP3K4、DmMAP3K7、DmMAP3K11、DmMAP4K5的转录表达水平与对照组相比，极显著上调。结果提示，MAPK基因家族成员与大型溞对nCeO2的胁迫应答调控相关。本研究为评估纳米材料对水生生物的潜在毒性提供科学参考。
The MAPK gene family is widely involved in various life processes in the cells of living organisms and is associated with the response of organisms to environmental stresses. However, the mechanism of MAPK action in Daphnia magna exposed to Nano cerium dioxide (nCeO2) is still unclear. Herein, we used bioinformatics methods such as BLAST, HMM and SMART to identify MAPK gene family members in the Daphnia magna genome, and analyzed the physicochemical properties, chromosomal localization, structural domains, conserved motifs, and phylogenetic relationships of individual MAPK genes in Daphnia magna. The MAPK gene family encodes proteins with sequence lengths of 361~1546 aa, molecular weights of 38.93~77.02 kDa, and isoelectric points in the range of 5.3~9.51. Twelve MAPK proteins have the S_TKc structural domain and four conserved motifs. Phylogenetic analysis showed that the 14 MAPK genes of Daphnia magna belonged to four subfamilies, and the gene sequences were highly conserved during evolution. The expression of seven MAPK genes, DmMAPK-p38b, DmMAPK4, DmMAP2K4, DmMAP3K4, DmMAP3K7, DmMAP3K11, and DmMAP4K5, was highly significantly up-regulated in Daphnia magna compared with the control group after 48 h of exposure to 50 and 150 mg/L nCeO2. The results suggest that members of the MAPK gene family are associated with the stress response of Daphnia magna to nCeO2 exposure. This study provides a scientific reference for assessing the potential toxicity of nanomaterials to aquatic organisms.
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