Abstract
Cnidarians are one of the oldest known animal lineages (ca. 700 million years), with a unique envenomation apparatus to deliver a potent mixture of peptides and proteins. Some peptide toxins from cnidarian venom have proven therapeutic potential. Here, we use a transcriptomic/proteomic strategy to identify sequences with similarity to known venom protein families in the tentacles of the endemic Australian ‘speckled anemone’ (Oulactis sp.). Illumina RNASeq data were assembled de novo. Annotated sequences in the library were verified by cross-referencing individuals’ transcriptomes or protein expression evidence from LC-MS/MS data. Sequences include pore-forming toxins, phospholipases, peptidases, neurotoxins (sodium and potassium channel modulators), cysteine-rich secretory proteins and defensins (antimicrobial peptides). Fewer than 4% of the sequences in the library occurred across the three individuals examined, demonstrating high sequence variability of an individual’s arsenal. We searched for actinoporins in Oulactis sp. to assess sequence similarity to the only described toxins (OR-A and -G) for this genus and examined the domain architecture of venom-related peptides and proteins. The novel putative actinoporin of Oulactis sp. has a greater similarity to other species in the Actiniidae family than to O. orientalis. Venom-related sequences have an architecture that occurs in single, repeat or multi-domain combinations of venom-related (e.g. ShK-like) and non-venom (e.g. whey acid protein) domains. This study has produced the first transcriptomes for an endemic Australian sea anemone species and the genus Oulactis, while identifying nearly 400 novel venom-related peptides and proteins for future structural and functional analyses and venom evolution studies.
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Acknowledgments
We acknowledge staff at the Monash Biomedical Proteomics Facility and Dr. Nathan Croft for proteomic assistance; Ms. Esperanza Rivera-de-Torre for supplying S. helianthus actinoporin sequences and Dr. Andrew Hugall, Ms. Fiona Boyle and Dr. Karin Luna-Ramirez for the helpful discussions. Computational resources were supported by the R@CMon/Monash Node of the NeCTAR Research Cloud, an initiative of the Australian Government’s Super Science Scheme and the Education Investment Fund.
Funding
This project was funded in part by ARC linkage grant LP150100621. M.L.M. received an Australian Government Research Training Program Scholarship, Monash Medicinal Chemistry Faculty Scholarship and Monash University-Museum Victoria Scholarship top-up. R.S.N., A.T.P. and A.W.P. received fellowship support from the Australian National Health and Medical Research Council.
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All authors contributed to the study conception and design. Material collection was performed by M.L.M. and R.A.V.M. and material preparation for transcriptome studies by M.L.M. Material preparation for mass spectrometry experiments was performed by M.L.M. and R.A.V.M.. Data collection was performed by M.L.M. and G.Q.T. and data analysis by M.L.M. All authors read and approved the final manuscript.
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All animal studies were carried out in accordance with the Victorian Fisheries Authority collection permit and Monash University Animal Ethics Regulations, Guidelines and Codes.
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Associated supplementary material may be found in the online version of this article and in the supplementary material and the online data set. Supplementary data files 1 through 6 are lodged as an online dataset in the data repository zenodo.org (https://doi.org/10.5281/zenodo.2561554) and Supplementary data files 7–9 may be found in the online version of this article;
Data file 1. Oulactis_sp_tentacle_Indv1_quant.sf
Data file 2. Oulactis_sp_tentacle_Indv2_quant.sf
Data file 3. Oulactis_sp_tentacle_Indv3_quant.sf
Data file 4. Indv1_Oulactis_sp_Trinotate_Report_1e-10.xlsx
Data file 5. Indv2_Oulactis_sp_Trinotate_Report_1e-10.xlsx
Data file 6. Indv3_Oulactis_sp_Trinotate_Report_1e-10.xlsx
Data file 7. VenomComponentLibrary.xlsx
Data file 8. Actinoporin phylogeny sequence alignment
Data file 9. Sequences of domain occurrence examples.xlsx
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Mitchell, M.L., Tonkin-Hill, G.Q., Morales, R.A.V. et al. Tentacle Transcriptomes of the Speckled Anemone (Actiniaria: Actiniidae: Oulactis sp.): Venom-Related Components and Their Domain Structure. Mar Biotechnol 22, 207–219 (2020). https://doi.org/10.1007/s10126-020-09945-8
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DOI: https://doi.org/10.1007/s10126-020-09945-8