Peroxiredoxin 6 is essential for the posttranslational activation of xanthine dehydrogenase in the uric acid synthesis of Bombyx mori.

Abstract

We identified a novel mutant of Bombyx mori, designated as male-absent oily (genetic symbol: om). The larval integument of this mutant is translucent due to a lack of uric acid in the integument. This mutation is Z-linked, and as mutant females are infertile, it is impossible to obtain om homozygous males. Using positional cloning combined with RNA-seq analysis, we identified a 1-bp deletion in the B. mori peroxiredoxin 6 (BmPrx6) gene. CRISPR/Cas9 knockout of BmPrx6 resulted in a translucent larval integument, indicating BmPrx6 as the causative gene for the om locus. Xanthine dehydrogenase (XDH)/xanthine oxidase (XO) is a key enzyme for uric acid synthesis. Injection of bovine XO into om mutants rescued the translucent phenotype, indicating that om is a mutant with defective XDH activity. To investigate XDH in B. mori, we generated a FLAG-tagged XDH gene using the CRISPR/Cas9 knock-in approach. Western blot analysis of XDH in om mutants revealed that BmPrx6 is crucial for the posttranslational activation of XDH. The role of BmPrx6 in regulating XDH activity is discussed.