Zeitschrift für Aquakulturforschung und -entwicklung

Abstrakt

Nitazoxanide Activating keap1a/Nrf2 Signaling Pathway is Regulated by Cul3 of Criataria plicata

Wuting Lu, Feixiang Su, Fanhua Yang, Jinhua An, Baoqing Hu, Shaoqing Jian, Gang Yang*, Chungen Wen*

Cul3 regulates the ubiquitination and degradation of Nrf2 in Nrf2/keap1a pathway under normal conditions. In aquatic invertebrates, the related mechanism of activating oxidative stress in mussels was studied by stimulating Cristatra plicata with Nitazoxanide (NTZ). The full-length Cul3 gene of C. plicata, designated CpCul3 was cloned. The comparative sequence analysis of CpCul3 gene and its homologous genes showed that CpCul3 gene was highly similar to the homologous genes of other species in amino acid sequence. The amino acid sequence of CpCul3 is 78%-98% similar to that of other invertebrates. In the phylogenetic tree, CpCul3 gene is more closely related to the Cul3 gene in other bivalves. After NTZ stimulation, the expression level of CpCul3 mRNA increased in hepatopancreas of the C. plicata. The protein expression level of CpCul3 was detected by utilizing CpCul3 specific antibody, and the expression level also increased. The results of subcellular localization of CpCul3 in HEK293T indicated that CpCul3 could co-locate with mitochondria, lysosomes, and endoplasmic reticulum, and the colocalization with mitochondria is more obvious. The results indicated that CpCul3 may plays an important role in regulating oxidative stress. The CoIP experiment proved that CpCul3 could promote the ubiquitination degradation of CpNrf2. Lastly, CpNrf2 could be degraded by K48-mediated ubiquitination.