Elucidating the transcriptional and post-translational systems that regulate snake venom assembly and that control autolysis and self-toxicity

阐明调节蛇毒组装并控制自溶和自毒的转录和翻译后系统

• 批准号: BB/F012675/1
• 负责人: Robert Harrison
• 金额: $ 40.13万
• 依托单位: Liverpool School of Tropical Medicine
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2008
• 资助国家: 英国
• 起止时间: 2008 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FF012675%2F1
• 关键词: Elucidating; transcriptional; post; translational; systems

Viper venom consists of a mixture of nature's most tissue-destructive proteins. These enzymes cause rapid and catastrophic cardiovascular collapse in envenomed rodent prey and, in humans, life-threatening bleeding and non-clotting blood. Yet while this lethal toxin cocktail is stored in the snake's venom gland it is somehow controlled so that the snakes' own tissues are unaffected. This extraordinary biological phenomenon is very poorly understood but is clearly central to the successful evolutionary development of venom as a predation and defensive/offensive survival strategy for venomous snakes. The control of viper protein toxicity has been very little studied because the appropriate gene and protein identification technologies have only recently become available. Previously, using basic biochemical techniques we and others have shown that one group of venom enzymes are kept in a quiescent state until activated by other, unknown, venom enzymes in an equally unknown time frame. It is also believed venom contains inhibitors to three key viper venom toxins but this has yet to be experimentally verified. We have recently developed expertise and resources in the application of new high throughput methods to identify new viper venom genes. Here, in the first study of its kind, we aim to provide the first holistic view of how sets of toxin and toxin-regulating venom gland genes are orchestrated during venom synthesis. Assisted by new gene and protein quantification technologies, we will first identify the genes and their protein products that constitute a model viper venom. Next, we will monitor the changes in the levels of these genes and proteins at several intervals spanning the time from when the snake ejects venom to the time it re-synthesises a fully competent arsenal of new venom. Using computational techniques, we will distinguish between toxins and proteins predicted to regulate their toxic activity. We will use molecular techniques to identify whether the genetic machinery controlling the production of (i) the toxins and (ii) the toxin-regulatory proteins, are co-ordinated to ensure that the regulatory proteins are produced before the respective toxin. The results of this study will greatly improve our understanding of the dynamics and the fundamental biological principles regulating venom assembly and venom toxicity.

毒蛇毒液由自然界最具组织破坏性的蛋白质混合物组成。这些酶会导致有毒的啮齿动物猎物迅速发生灾难性的心血管衰竭，而对于人类来说，会导致危及生命的出血和不凝固的血液。然而，虽然这种致命的毒素混合物储存在蛇的毒腺中，但它以某种方式受到控制，因此蛇自身的组织不受影响。人们对这种非凡的生物现象知之甚少，但显然对于毒液作为毒蛇的捕食和防御/进攻生存策略的成功进化发展至关重要。由于适当的基因和蛋白质鉴定技术最近才出现，因此对蝰蛇蛋白质毒性控制的研究很少。此前，我们和其他人使用基本的生化技术表明，一组毒液酶保持静止状态，直到在同样未知的时间范围内被其他未知的毒液酶激活。人们还认为毒液含有三种主要毒蛇毒毒素的抑制剂，但这尚未得到实验验证。我们最近开发了应用新的高通量方法来识别新的毒蛇毒液基因的专业知识和资源。在这里，在此类研究中，我们的目标是提供关于毒素和毒素调节毒腺基因组在毒液合成过程中如何协调的第一个整体视图。在新的基因和蛋白质定量技术的帮助下，我们将首先鉴定构成毒蛇毒液模型的基因及其蛋白质产物。接下来，我们将在蛇喷射毒液到重新合成完全有效的新毒液期间的几个时间间隔内监测这些基因和蛋白质水平的变化。使用计算技术，我们将区分毒素和预测调节其毒性活性的蛋白质。我们将使用分子技术来确定控制（i）毒素和（ii）毒素调节蛋白产生的遗传机制是否协调，以确保调节蛋白在相应毒素之前产生。这项研究的结果将极大地提高我们对调节毒液组装和毒液毒性的动力学和基本生物学原理的理解。

项目成果

Comparative venom gland transcriptome surveys of the saw-scaled vipers (Viperidae: Echis) reveal substantial intra-family gene diversity and novel venom transcripts.

比较毒腺的比较转录组调查（Viperidae：echis）显示了家族内基因多样性和新型毒液转录本。

• DOI: 10.1186/1471-2164-10-564
• 发表时间: 2009-11-30
• 期刊: BMC genomics
• 影响因子: 4.4
• 作者: Casewell NR;Harrison RA;Wüster W;Wagstaff SC
• 通讯作者: Wagstaff SC

Pre-clinical assays predict pan-African Echis viper efficacy for a species-specific antivenom.

• DOI: 10.1371/journal.pntd.0000851
• 发表时间: 2010-10-26
• 期刊: PLoS neglected tropical diseases
• 影响因子: 3.8
• 作者: Casewell NR;Cook DA;Wagstaff SC;Nasidi A;Durfa N;Wüster W;Harrison RA
• 通讯作者: Harrison RA