Key Laboratory of Tropical Biological Resources of Ministry of Education, Hainan University, Haikou, Hainan, China.
Laboratory of Tropical Veterinary Medicine and Vector Biology, Institute of Tropical Agriculture and Forestry, Hainan University, Haikou, Hainan, China.
PLoS One. 2018 Mar 19;13(3):e0194499. doi: 10.1371/journal.pone.0194499. eCollection 2018.
Mosquitoes transmit a number of diseases in animals and humans, including Dengue, Chikungunya and Zika viruses that affect millions of people each year. Controlling the disease-transmitting mosquitoes has proven to be a successful strategy to reduce the viruses transmission. Polyamines are required for the life cycle of the RNA viruses, Chikungunya virus and Zika virus, and a depletion of spermidine and spermine in the host via induction of spermine N-acetyltransferase restricts their replication. Spermine N-acetyltransferase is a key catabolic enzyme in the polyamine pathway, however there is no information of the enzyme identification in any insects. Aliphatic polyamines play a fundamental role in tissue growth and development in organisms. They are acetylated by spermidine/spermine N1-acetyltransferase (SAT). In this study we provided a molecular and biochemical identification of SAT from Aedes aegypti mosquitoes. Screening of purified recombinant proteins against polyamines established that aaNAT5b, named previously based on sequence similarity with identified aaNAT1 in insects, is active to spermine and spermidine. A crystal structure was determined and used in molecular docking in this study. Key residues were identified to be involved in spermine binding using molecular docking and simulation. In addition, SAT transcript was down regulated by blood feeding using a real time PCR test. Based on its substrate profile and transcriptional levels after blood feeding, together with previous reports for polyamines required in arboviruses replication, SAT might be potentially used as a target to control arboviruses with human interference.
蚊子在动物和人类中传播多种疾病,包括登革热、基孔肯雅热和寨卡病毒,这些病毒每年影响数百万人。控制传播疾病的蚊子已被证明是减少病毒传播的成功策略。多胺是 RNA 病毒(如基孔肯雅热病毒和寨卡病毒)生命周期所必需的,通过诱导精脒 N-乙酰转移酶使宿主中的精脒和精胺耗竭,限制了它们的复制。精脒 N-乙酰转移酶是多胺途径中的关键分解代谢酶,但在任何昆虫中都没有该酶的鉴定信息。脂肪族多胺在生物体的组织生长和发育中起着基本作用。它们被精脒/精胺 N1-乙酰转移酶(SAT)乙酰化。在这项研究中,我们从埃及伊蚊中提供了 SAT 的分子和生化鉴定。针对多胺对纯化重组蛋白的筛选表明,以前根据与昆虫中鉴定的 aaNAT1 的序列相似性命名的 aaNAT5b 对精脒和精胺具有活性。本研究确定了一个晶体结构,并在分子对接中使用了该结构。通过分子对接和模拟确定了关键残基参与精脒结合。此外,SAT 转录本通过实时 PCR 测试在吸血后下调。基于其底物谱和吸血后的转录水平,以及先前关于多胺在虫媒病毒复制中的作用的报告,SAT 可能可作为人类干预控制虫媒病毒的潜在靶点。