{"title":"将双链 RNA 分解为 20-nt 小 RNA 的森蚕蛾 Dicer-2 的生化特征。","authors":"Midori Tabara, Mayuko Harada, Kazunori Kuriyama, Takuma Sakamoto, Atsushi Takeda, Toshiyuki Fukuhara, Hiroko Tabunoki","doi":"10.1002/arch.22118","DOIUrl":null,"url":null,"abstract":"<p>We detected enzymatic activity that generates 20-nucleotide (nt) RNA from double-stranded RNAs (dsRNAs) in crude extracts prepared from various silkworm (<i>Bombyx mori</i>) organs. The result using knocked-down cultured cells indicated that this dicing activity originated from <i>B. mori</i> Dicer-2 (BmDcr2). Biochemical analyses revealed that BmDcr2 preferentially cleaves 5′-phosphorylated dsRNAs at the 20-nt site—counted from the 5′-phosphorylated end—and required ATP and magnesium ions for the dicing reaction. This is the first report of the biochemical characterization of Dicer-2 in lepidopteran insects. This enzymatic property of BmDcr2 in vitro is consistent with the in vivo small interfering RNA profile in virus-infected silkworm cells.</p>","PeriodicalId":8281,"journal":{"name":"Archives of Insect Biochemistry and Physiology","volume":null,"pages":null},"PeriodicalIF":1.5000,"publicationDate":"2024-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Biochemical characterization of Bombyx mori Dicer-2 that dices double-stranded RNAs into 20-nt small RNA\",\"authors\":\"Midori Tabara, Mayuko Harada, Kazunori Kuriyama, Takuma Sakamoto, Atsushi Takeda, Toshiyuki Fukuhara, Hiroko Tabunoki\",\"doi\":\"10.1002/arch.22118\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>We detected enzymatic activity that generates 20-nucleotide (nt) RNA from double-stranded RNAs (dsRNAs) in crude extracts prepared from various silkworm (<i>Bombyx mori</i>) organs. The result using knocked-down cultured cells indicated that this dicing activity originated from <i>B. mori</i> Dicer-2 (BmDcr2). Biochemical analyses revealed that BmDcr2 preferentially cleaves 5′-phosphorylated dsRNAs at the 20-nt site—counted from the 5′-phosphorylated end—and required ATP and magnesium ions for the dicing reaction. This is the first report of the biochemical characterization of Dicer-2 in lepidopteran insects. This enzymatic property of BmDcr2 in vitro is consistent with the in vivo small interfering RNA profile in virus-infected silkworm cells.</p>\",\"PeriodicalId\":8281,\"journal\":{\"name\":\"Archives of Insect Biochemistry and Physiology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":1.5000,\"publicationDate\":\"2024-05-07\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Archives of Insect Biochemistry and Physiology\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/arch.22118\",\"RegionNum\":4,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Archives of Insect Biochemistry and Physiology","FirstCategoryId":"97","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/arch.22118","RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
我们在从各种家蚕（Bombyx mori）器官制备的粗提取物中检测到了从双链 RNA（dsRNA）生成 20 核苷酸（nt）RNA 的酶活性。使用基因敲除培养细胞的结果表明，这种切割活性来自于 B. mori Dicer-2（BmDcr2）。生化分析表明，BmDcr2 优先在 20-nt 位点（从 5'- 磷酸化末端开始计算）切割 5'- 磷酸化 dsRNA，切割反应需要 ATP 和镁离子。这是首次报道鳞翅目昆虫中 Dicer-2 的生化特性。BmDcr2 在体外的这种酶特性与体内病毒感染蚕细胞中的小干扰 RNA 特征是一致的。
Biochemical characterization of Bombyx mori Dicer-2 that dices double-stranded RNAs into 20-nt small RNA
We detected enzymatic activity that generates 20-nucleotide (nt) RNA from double-stranded RNAs (dsRNAs) in crude extracts prepared from various silkworm (Bombyx mori) organs. The result using knocked-down cultured cells indicated that this dicing activity originated from B. mori Dicer-2 (BmDcr2). Biochemical analyses revealed that BmDcr2 preferentially cleaves 5′-phosphorylated dsRNAs at the 20-nt site—counted from the 5′-phosphorylated end—and required ATP and magnesium ions for the dicing reaction. This is the first report of the biochemical characterization of Dicer-2 in lepidopteran insects. This enzymatic property of BmDcr2 in vitro is consistent with the in vivo small interfering RNA profile in virus-infected silkworm cells.
期刊介绍：
Archives of Insect Biochemistry and Physiology is an international journal that publishes articles in English that are of interest to insect biochemists and physiologists. Generally these articles will be in, or related to, one of the following subject areas: Behavior, Bioinformatics, Carbohydrates, Cell Line Development, Cell Signalling, Development, Drug Discovery, Endocrinology, Enzymes, Lipids, Molecular Biology, Neurobiology, Nucleic Acids, Nutrition, Peptides, Pharmacology, Pollinators, Proteins, Toxicology. Archives will publish only original articles. Articles that are confirmatory in nature or deal with analytical methods previously described will not be accepted.
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