Laboratoire d'Ingénierie des Anticorps pour la Santé (LIAS), CEA/DSV/iBiTec-s/SPI, CEA Saclay, Bt. 152, 91191 Gif-sur-Yvette Cedex, France.
Toxicon. 2012 Jan;59(1):34-46. doi: 10.1016/j.toxicon.2011.10.001. Epub 2011 Nov 6.
Although cone snail venoms have been intensively investigated in the past few decades, little is known about the whole conopeptide and protein content in venom ducts, especially at the transcriptomic level. If most of the previous studies focusing on a limited number of sequences have contributed to a better understanding of conopeptide superfamilies, they did not give access to a complete panorama of a whole venom duct. Additionally, rare transcripts were usually not identified due to sampling effect. This work presents the data and analysis of a large number of sequences obtained from high throughput 454 sequencing technology using venom ducts of Conus consors, an Indo-Pacific living piscivorous cone snail. A total of 213,561 Expressed Sequence Tags (ESTs) with an average read length of 218 base pairs (bp) have been obtained. These reads were assembled into 65,536 contiguous DNA sequences (contigs) then into 5039 clusters. The data revealed 11 conopeptide superfamilies representing a total of 53 new isoforms (full length or nearly full-length sequences). Considerable isoform diversity and major differences in transcription level could be noted between superfamilies. A, O and M superfamilies are the most diverse. The A family isoforms account for more than 70% of the conopeptide cocktail (considering all ESTs before clustering step). In addition to traditional superfamilies and families, minor transcripts including both cysteine free and cysteine-rich peptides could be detected, some of them figuring new clades of conopeptides. Finally, several sets of transcripts corresponding to proteins commonly recruited in venom function could be identified for the first time in cone snail venom duct. This work provides one of the first large-scale EST project for a cone snail venom duct using next-generation sequencing, allowing a detailed overview of the venom duct transcripts. This leads to an expanded definition of the overall cone snail venom duct transcriptomic activity, which goes beyond the cysteine-rich conopeptides. For instance, this study enabled to detect proteins involved in common post-translational maturation and folding, and to reveal compounds classically involved in hemolysis and mechanical penetration of the venom into the prey. Further comparison with proteomic and genomic data will lead to a better understanding of conopeptides diversity and the underlying mechanisms involved in conopeptide evolution.
尽管过去几十年人们对芋螺毒液进行了深入研究,但对于毒液管中的整个芋螺肽和蛋白质含量却知之甚少,尤其是在转录组水平上。如果之前的大多数研究集中在有限数量的序列上,这有助于更好地理解芋螺肽超家族,但它们无法全面了解整个毒液管。此外,由于采样效应,通常无法识别罕见的转录本。本工作展示了使用来自印度-太平洋地区肉食性芋螺 Conus consors 的毒液管,通过高通量 454 测序技术获得的大量序列的数据和分析。共获得了 213561 条平均读长为 218 个碱基对(bp)的表达序列标签(EST)。这些读段被组装成 65536 条连续的 DNA 序列(contigs),然后进一步聚类成 5039 个簇。这些数据揭示了 11 种芋螺肽超家族,代表了总共 53 种新的同工型(全长或近全长序列)。不同超家族之间可以观察到显著的同工型多样性和转录水平的主要差异。A、O 和 M 超家族是最具多样性的。A 家族同工型占芋螺肽鸡尾酒的 70%以上(考虑聚类前的所有 EST)。除了传统的超家族和家族之外,还可以检测到包括无半胱氨酸和富含半胱氨酸的肽在内的少量转录本,其中一些构成了新的芋螺肽分支。最后,首次在芋螺毒液管中鉴定出与毒液功能相关的几种蛋白质转录本。这项工作提供了第一个使用下一代测序的芋螺毒液管的大规模 EST 项目之一,允许对毒液管转录本进行详细概述。这导致了对整体芋螺毒液管转录组活性的扩展定义,超出了富含半胱氨酸的芋螺肽的范围。例如,这项研究能够检测到参与常见翻译后成熟和折叠的蛋白质,并揭示了经典地参与毒液进入猎物的溶血和机械渗透的化合物。与蛋白质组学和基因组学数据的进一步比较将有助于更好地理解芋螺肽的多样性以及芋螺肽进化背后的机制。