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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Laccase 2 is the phenoloxidase gene required for beetle cuticle tanning
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Laccase 2 is the phenoloxidase gene required for beetle cuticle tanning

机译：漆酶2是甲虫角质层鞣制所需的酚氧化酶基因

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摘要

Cuticle tanning (or sclerotization and pigmentation) in invertebrates involves the oxidative conjugation of proteins, which renders them insoluble and hardens and darkens the color of the exoskeleton. Two kinds of phenoloxidases, laccase and tyrosinase, have been proposed to participate in tanning, but proof of the true identity of the enzyme(s) responsible for this process has been elusive. We report the cloning of cDNAs for laccases and tyrosi-nases from the red flour beetle, Tribolium castaneum, as well as their developmental patterns of expression. To test for the involvement of these types of enzymes in cuticle tanning, we performed RNA interference experiments to decrease the levels of individual phenoloxidases. Normal phenotypes were obtained after dsRNA-mediated transcript depletion for all phenoloxidases tested, with the exception of laccase 2. Insects injected with dsRNA for the laccase 2 gene failed to tan, were soft-bodied and deformed, and subsequently died in a dsRNA dose-dependent fashion. The results presented here support the hypothesis that two isoforms of laccase 2 generated by alternative splicing catalyze larval, pupal, and adult cuticle tanning in Tribolium.
机译：无脊椎动物的角质层鞣制（或硬化和色素沉着）涉及蛋白质的氧化共轭，使蛋白质不溶并硬化和加深外骨骼的颜色。已经提出了两种酚氧化酶，漆酶和酪氨酸酶参与鞣制，但是难以证明负责该过程的酶的真实身份。我们报告了从红色面粉甲虫Tribolium castaneum的漆酶和酪氨酸酶cDNA的克隆及其表达的发育模式。为了测试这些类型的酶在角质层鞣革中的参与，我们进行了RNA干扰实验以降低单个酚氧化酶的水平。 dsRNA介导的所有酚氧化酶的转录本耗竭后均获得正常表型，漆酶2除外。为漆酶2基因注入dsRNA的昆虫无法晒黑，变软并变形，随后在dsRNA剂量下死亡。依赖时尚。此处提出的结果支持以下假设：通过交替剪接生成的漆酶2的两个同工型催化Tribolium中的幼虫，p和成年表皮鞣制。

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