Bhar Subhradeep, Prajapati Dilip V, Gonzalez Melisa S, Yoon Chi-Su, Mai Kevin, Bailey Laura S, Basso Kari B, Butcher Rebecca A
Department of Chemistry, University of Florida, Gainesville, Florida 32611, United States.
ACS Chem Biol. 2025 Jun 20;20(6):1298-1308. doi: 10.1021/acschembio.5c00126. Epub 2025 Jun 5.
The nematode produces a large family of ascaroside pheromones, which it uses in chemical communication to coordinate the development and behavior of the population. The acyl-CoA oxidase (ACOX) enzymes, which catalyze the first rate-limiting step in peroxisomal β-oxidation, act as gatekeepers for the biosynthesis of ascarosides with specific side-chain lengths. By performing unbiased comparative metabolomics on , , , , and mutant worms and double mutant worms, we provide a comprehensive view of the different roles of these enzymes in ascaroside biosynthesis and implicate them in a number of additional biosynthetic pathways. Our data show that and are required for the biosynthesis of a broad range of medium- and long-chain ascarosides, while , , and specialize in ascarosides with specific side-chain lengths. Specific mutants accumulate a variety of modified ascarosides that are likely shunt products. Furthermore, we show that and , but not other genes, are required for the biosynthesis of a specific subset of -acylethanolamines (NAEs), many of which have hydroxyl groups at specific positions in their fatty acyl side chains. Through stable-isotope labeling, feeding experiments, and chemical synthesis, we characterize the structures of these NAEs and show that their fatty acyl groups are derived from both bacteria and nematode sources. One of the most strongly -dependent NAEs that has a β-hydroxy fatty acyl group is attractive to at attomolar concentrations, whereas a closely related NAE with a γ-hydroxy fatty acyl group is not, indicating that a subset of secreted NAEs may influence worm behavior.
这种线虫产生一大类ascaroside信息素，用于化学通讯以协调群体的发育和行为。酰基辅酶A氧化酶（ACOX）催化过氧化物酶体β-氧化的第一步限速反应，在具有特定侧链长度的ascarosides生物合成中起把关作用。通过对野生型、acox-1、acox-2、acox-3和acox-4突变线虫以及acox-1;acox-2双突变线虫进行无偏向比较代谢组学研究，我们全面了解了这些酶在ascaroside生物合成中的不同作用，并发现它们参与了许多其他生物合成途径。我们的数据表明，ACOX-1和ACOX-2是多种中链和长链ascarosides生物合成所必需的，而ACOX-3、ACOX-4和ACOX-5专门参与具有特定侧链长度的ascarosides的合成。特定的acox突变体积累了多种可能是分流产物的修饰ascarosides。此外，我们表明，ACOX-1和ACOX-2，但不是其他ACOX基因，是特定子集的N-酰基乙醇胺（NAEs）生物合成所必需的，其中许多在其脂肪酰基侧链的特定位置具有羟基。通过稳定同位素标记、喂食实验和化学合成，我们表征了这些NAEs的结构，并表明它们的脂肪酰基既来源于细菌也来源于线虫。其中一种对ACOX-1依赖性最强且具有β-羟基脂肪酰基的NAE在阿托摩尔浓度下对秀丽隐杆线虫有吸引力，而一种具有γ-羟基脂肪酰基的密切相关的NAE则没有，这表明分泌的NAEs子集中的一些可能会影响线虫的行为。
