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

IF 3.2 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology Pub Date : 2025-01-29 DOI:10.1016/j.ibmb.2025.104264

Tsuguru Fujii , Masato Hino , Toshiaki Fujimoto , Kohei Kakino , Yu Kaneko , Hiroaki Abe , Jae Man Lee , Takahiro Kusakabe , Toru Shimada

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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.

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家蚕尿酸合成过程中，过氧化物还氧蛋白6对黄嘌呤脱氢酶的翻译后激活至关重要。

我们鉴定了一种新的家蚕突变体，命名为雄性无油（遗传符号：om）。由于缺乏尿酸，这种突变体的幼虫外壳是半透明的。这种突变是z连锁的，由于突变的雌性是不育的，所以不可能获得纯合的雄性。利用定位克隆与RNA-seq分析相结合的方法，我们在家蚕过氧化物还氧蛋白6 （BmPrx6）基因中发现了一个1 bp的缺失。CRISPR/Cas9敲除BmPrx6导致半透明的幼虫被膜，表明BmPrx6是om位点的致病基因。黄嘌呤脱氢酶(XDH)/黄嘌呤氧化酶（XO）是尿酸合成的关键酶。将牛XO注射到om突变体中，恢复了半透明表型，表明om是XDH活性缺陷的突变体。为了研究家蚕中的XDH，我们使用CRISPR/Cas9敲入方法生成了一个flag标记的XDH基因。XDH的Western blot分析显示，BmPrx6对XDH的翻译后激活至关重要。讨论了BmPrx6在调节XDH活性中的作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Insect Biochemistry and Molecular Biology 生物-昆虫学

CiteScore

7.40

自引率

5.30%

发文量

105

审稿时长

40 days

期刊介绍： This international journal publishes original contributions and mini-reviews in the fields of insect biochemistry and insect molecular biology. Main areas of interest are neurochemistry, hormone and pheromone biochemistry, enzymes and metabolism, hormone action and gene regulation, gene characterization and structure, pharmacology, immunology and cell and tissue culture. Papers on the biochemistry and molecular biology of other groups of arthropods are published if of general interest to the readership. Technique papers will be considered for publication if they significantly advance the field of insect biochemistry and molecular biology in the opinion of the Editors and Editorial Board.