Molecular characteristics and immune function of ubiquitin C-terminal hydrolase-L3 in Macrobrachium nipponense.

ABSTRACT

Ubiquitin C-terminal hydrolase-L3 (UCHL3) is a deubiquitinating enzyme involved in the repair mechanism of homologous recombinations of DNA double strand breaks (DBS). However, the role of UCHL3 in crustacean immune regulation has not been studied. In this experiment, we cloned and analyzed the expression profile of the UCHL3 gene from Macrobrachium nipponense (MnUCHL3). The obtained full-length cDNA of the MnUCHL3 transcript was 1192 bp, and it had a 687 bp open reading frame encoding a 228 amino acid protein, and the structure of UCHL3 is highly similar to that of other invertebrates. Real-time PCR results indicated that MnUCHL3 was expressed in all detected tissues, with the highest expression levels in the hepatopancreas, and the expression of MnUCHL3 in the gill and hepatopancreas was downregulated to different degrees within 48 h after the infection of viruses and bacteria. Furthermore, knockdown of MnUCHL3 expression by double-stranded RNA (dsRNA) injection in Aeromonas hydrophila-infected prawns increased prawn mortality and bacterial growth. In addition, overexpression of MnUCHL3 in HEK293T cells in vitro suggested that MnUCHL3 could activate the NF-κB signal path and the expression levels of NF-κB signaling cascade members and AMPs, exhibiting remarkable downregulation in the MnUCHL3-silenced group. The above experimental conclusions revealed that UCHL3 gene might be involved in the innate immune response to bacterial infection by regulating the synthesis of a series of AMPs, and these results might provide new insights into UCHL3 in invertebrates.