Characterization of an agmatine N-acetyltransferase from Bactrocera dorsalis that modulates ovary development

• Published in: Insect biochemistry and molecular biology 2024-07, Vol.170, p.104130, Article 104130
• Main Authors: Teng, Fei-yue, Feng, Ji-mei, Ma, Fu-cai, Wang, Zhuo-xin, Lu, Yong-yue, Qi, Yi-xiang
• Format: Article
• Language: English
• Subjects: Agmatine; Bactrocera dorsalis; N-acetyltransferase; Ovary development; Steady-state kinetics
• ISSN: 0965-1748; 1879-0240; 1879-0240
• DOI: 10.1016/j.ibmb.2024.104130

Agmatine N-acetyltransferase (AgmNAT), which catalyzes the formation of N-acetylagmatine from acetyl-CoA and agmatine, is a member of the GCN5-related N-acetyltransferase family. So far, knowledge of the physiological roles of AgmNAT in insects is limited. Here, we identified one gene encoding protein homologous to that of Drosophila AgmNAT using sequence information from an activity-verified Drosophila AgmNAT in a BLAST search of the Bactrocera dorsalis genome. We expressed and purified B. dorsalis AgmNAT in Escherichia coli and used the purified enzyme to define the substrate specificity for acyl-CoA and amine substrates. Our application of the screening strategy to BdorAgmNAT led to the identification of agmatine as the best amine substrate for this enzyme, with the highest kcat/Km value. We successfully obtained a BdorAgmNAT knockout strain based on a wild-type strain (WT) using the CRISPR/Cas9 technique. The ovary development of the BdorAgmNAT knockout mutants was delayed for 10 days compared with the WT specimens. Moreover, mutants had a much smaller mature ovary size and laid far fewer eggs than WT. Loss of function of BdorAgmNAT caused by RNAi with mature WT females did not affect their fecundity. These findings indicate that BdorAgmNAT is critical for oogenesis. Our data provide the first evidence for AgmNAT in regulating ovary development. [Display omitted] •Enzymatic properties of the purified recombinant BdorAgmNAT was analyzed.•We generated a BdorAgmNAT knockout mutant using the CRISPR/Cas9 system.•BdorAgmNAT gene is critical for ovary development.