Acyl-CoA Oxidases Fine-Tune the Production of Ascaroside Pheromones with Specific Side Chain Lengths,ACS Chemical Biology

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Acyl-CoA Oxidases Fine-Tune the Production of Ascaroside Pheromones with Specific Side Chain Lengths
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2018-03-14 00:00:00 , DOI: 10.1021/acschembio.7b01021
Xinxing Zhang ₁ , Yuting Wang ₁ , David H. Perez ₁ , Rachel A. Jones Lipinski ₁ , Rebecca A. Butcher ₁

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Caenorhabditis elegans produces a complex mixture of ascaroside pheromones to control its development and behavior. Acyl-CoA oxidases, which participate in β-oxidation cycles that shorten the side chains of the ascarosides, regulate the mixture of pheromones produced. Here, we use CRISPR-Cas9 to make specific nonsense and missense mutations in acox genes and determine the effect of these mutations on ascaroside production in vivo. Ascaroside production in acox-1.1 deletion and nonsense strains, as well as a strain with a missense mutation in a catalytic residue, confirms the central importance of ACOX-1.1 in ascaroside biosynthesis and suggests that ACOX-1.1 functions in part by facilitating the activity of other acyl-CoA oxidases. Ascaroside production in an acox-1.1 strain with a missense mutation in an ATP-binding site at the ACOX-1.1 dimer interface suggests that ATP binding is important for the enzyme to function in ascaroside biosynthesis in vivo. Ascaroside production in strains with deletion, nonsense, and missense mutations in other acox genes demonstrates that ACOX-1.1 works with ACOX-1.3 in processing ascarosides with 7-carbon side chains, ACOX-1.4 in processing ascarosides with 9- and 11-carbon side chains, and ACOX-3 in processing ascarosides with 13- and 15-carbon side chains. It also shows that ACOX-1.2, but not ACOX-1.1, processes ascarosides with 5-carbon ω-side chains. By modeling the ACOX structures, we uncover characteristics of the enzyme active sites that govern substrate preferences. Our work demonstrates the role of specific acyl-CoA oxidases in controlling the length of ascaroside side chains and thus in determining the mixture of pheromones produced by C. elegans.

中文翻译：

酰基辅酶A氧化酶可微调具有特定侧链长度的A螨苷信息素的生产

秀丽隐杆线虫会产生一种复杂的mixture虫苷信息素混合物，以控制其发育和行为。参与缩短了A螨侧链的β-氧化循环的酰基辅酶A氧化酶可调节所产生的信息素的混合物。在这里，我们使用CRISPR-Cas9在acox基因中进行特定的无义和错义突变，并确定这些突变对体内a虫苷产生的影响。acox-1.1中的scar虫苷生产缺失和无意义菌株，以及在催化残基中具有错义突变的菌株，证实了ACOX-1.1在a虫苷生物合成中的重要作用，并表明ACOX-1.1的部分功能是通过促进其他酰基辅酶A氧化酶的活性来实现的。在acox-1.1菌株中产生的side虫苷在ACOX-1.1二聚体界面的ATP结合位点有错义突变，这表明ATP结合对于该酶在体内的a虫苷生物合成具有重要作用。在其他acox中缺失，无义和错义突变的菌株中产生scar螨甙基因证明ACOX-1.1与ACOX-1.3一起加工具有7个碳侧链的scar螨，ACOX-1.4与加工中具有9个碳和11个碳侧链的scar螨，而ACOX-3与ACOX-3一起加工具有13个碳和15个碳的侧scar螨。碳侧链。它也表明ACOX-1.2，而不是ACOX-1.1，加工带有5碳ω侧链的a螨。通过对ACOX结构进行建模，我们发现了控制底物偏好的酶活性位点的特征。我们的工作证明了特定的酰基辅酶A氧化酶在控制a糖侧链长度上的作用，从而在确定秀丽隐杆线虫产生的信息素混合物中的作用。

更新日期：2018-03-14

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