Transcription factor GATA5 enhances autophagy by modulating miR-2765 in Penaeus vannamei under ammonia nitrogen...

The transcription factor GATA is recognized as a key regulator of cell generation and differentiation, playing a significant regulatory role in immune responses triggered by both biotic and abiotic stress. However, the regulatory network of GATA in crustaceans remains poorly understood. In this study, we identified a GATA5-like protein, designated as PvGATA5, from Penaeus vannamei. The nucleic acid sequence length of PvGATA5 is 2203 bp, with an open reading frame (ORF) of 1134 bp and a predicted protein molecular weight of 40.77 kDa. PvGATA5 contains two conserved zinc finger domains characterized by the motif "Cys-X2-X17-Cys-X2-Cys" which are crucial for DNA binding. Phylogenetic tree analysis indicates that PvGATA5 is closely related to GATA4/6 in mammals. Additionally, PvGATA5 exhibits the highest expression in hemocytes and significantly enhances the hemocyte count and autophagy levels in shrimp under ammonia nitrogen stress, thereby improving shrimp survival rates. Furthermore, PvGATA5 is shown to recognize and bind to the "ACCCAGATAG" sequence in the promoter region of miR-2765, as demonstrated by dual luciferase reporter assays, ChIP-PCR and EMSA analysis, leading to the negative regulation of miR-2765 expression. Collectively, these findings suggest that PvGATA5 promotes autophagy and maintains hematopoietic homeostasis by negatively regulating miR-2765. This mechanism alleviates the stress damage caused by ammonia nitrogen in P. vannamei, thereby improving their survival rates. Our research provides a valuable foundation for further investigations into the functions and molecular mechanisms of GATA in invertebrates.