Acyl-CoA Oxidases Fine-Tune the Production of Ascaroside Pheromones with Specific Side Chain Lengths
Acyl-CoA Oxidases Fine-Tune the Production of Ascaroside Pheromones with Specific Side Chain Lengths
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DOI：
10.1021/acschembio.7b01021
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发表时间：
2018-04-01
影响因子：
4
通讯作者：
Butcher, Rebecca A.
中科院分区：
文献类型：
--
作者：
Zhang, Xinxing;Wang, Yuting;Butcher, Rebecca A.
秀丽隐杆线虫产生复杂的混合蛔虫苷信息素来控制其发育和行为。酰基辅酶a氧化酶参与β -氧化循环，缩短蛔虫苷的侧链，调节产生的信息素的混合物。在这里，我们使用CRISPR-Cas9在acox基因中制造特异性无义和错义突变，并确定这些突变对体内天冬苷产生的影响。在acox-1.1缺失菌株和无意义菌株以及催化残基错意突变菌株中，天红苷的产生证实了acox-1.1在天红苷生物合成中的核心重要性，并表明acox-1.1在一定程度上通过促进其他酰基辅酶a氧化酶的活性发挥作用。在acox-1.1二聚体界面的ATP结合位点发生错意突变的acox-1.1菌株产生了天冬苷，这表明ATP结合对于该酶在体内天冬苷生物合成中发挥作用是重要的。在其他辅酶基因缺失、无意义和错sense突变的菌株中产生的天牛苷表明，acox -1.1与acox -1.3一起加工7碳侧链的天牛苷，acox -1.4加工9碳和11碳侧链的天牛苷，acox -3加工13碳和15碳侧链的天牛苷。它还表明，ACOX-1.2，而不是ACOX-1.1，加工具有5碳ω -侧链的asasar苷。通过对ACOX结构进行建模，我们揭示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氧化酶在控制蛔虫侧链长度方面的作用，从而决定了秀丽隐杆线虫产生的信息素的混合物。
Caenorhabditis elegans produces a complex mixture of ascaroside pheromones to control its development and behavior. Acyl-CoA oxidases, which participate in beta-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 omega-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.