Characterization of a γ-aminobutyrate type A receptor-associated protein gene, which is involved in the response of Portunus trituberculatus to CO2 -induced ocean acidification

Abstract

The γ-aminobutyrate type A receptor-associated protein (GABARAP) is a ubiquitin-like modifier implicated in membrane trafficking and fusion events involving the γ-aminobutyrate type A receptor, autophagy and apoptosis. In this study, the gene encoding GABARAP was cloned from swimming crab Portunus trituberculatus (PtGABARAP) based on the expression sequence tag (EST). The full-length cDNA of 664 bp includes a 5′ untranslated region (UTR) of 87 bp, a 3′ UTR of 223 bp with a poly(A) tail, and an open reading frame (ORF) of 354 bp encoding a polypeptide of 117 amino acids with a predicted molecular weight of 13.96 kDa. The deduced amino acid sequence shares high similarity (93%–100%) with GABARAPs from other species and includes a conserved Atg8 domain. In a phylogenetic analysis PtGABARAP clustered with GABARAPs from other species, and more widely with other GABARAP family proteins. The impact of elevated ocean acidification (OA) on P. trituberculatus behaviours was investigated, and real-time RT-PCR revealed that PtGABARAP expression was up-regulated after OA exposure. Ocean acidification also caused crabs anxiety-like behaviours, like the shoal average speed increase, preference for dark environment (scototaxis) and fast exploration. The results indicated that GABARAP might be involved in the interactions of GABAA receptors and elevated-CO2 seawater.