Cloning and characterization of AMP-activated protein kinase genes in Daphnia pulex: Modulation of AMPK gene expression in response to polystyrene nanoparticles

Abstract

Energy metabolism is essential for almost all organisms. At the molecular level, adenosine monophosphate activated protein kinase (AMPK) plays a vital role in cellular energy homeostasis. Its molecular characterization in invertebrates, including Daphnia pulex, and the understanding of its role in response to environmental contaminants is limited. In this study, three subunits of AMPK (AMPKα, β, and γ) were cloned in D. pulex, and assigned the GenBank accession numbers MT536758, MT536759, and MT536760, respectively. Their full lengths were 2,000, 1,384, and 2594 bp, respectively, and contained open reading frames of 526, 274, and 580 amino acids, respectively. Bioinformatic analysis revealed that the three AMPK subunits all have features representative of the AMPK superfamily, and were correspondingly clustered with each orthologue branch. The three AMPK subunit genes, AMPKα, β, and γ, had the highest similarity to those of other organisms at 82%, 94%, and 71%, respectively. Nanoplastics significantly increased AMPKα expression, but decreased that of AMPKβ and γ. These results identified AMPKα, β, and γ in D. pulex, and showed that they all encode proteins with conserved functional domains. This study provides basic information on how three types of AMPK in aquatic organisms respond to environmental contaminants.