Animal venoms are complex biochemical cocktails capable of diverse and extremely potent physiological effects. Venoms are also highly prized as novel sources of pharmaceuticals and are essential for the development of lifesaving antivenoms. Despite their scientific and clinical significance, the molecular composition and functional activities of many biomedically important venoms remains poorly known. The long-term objective of this proposal is to understand the molecular complexity, function and evolution of black widow spider venom. Black widow spiders possess a highly neurotoxic venom that causes severe pain in humans. While black widow venom has played a critical role in the study of vertebrate neurotransmission, the molecular composition of this venom and its functional and biomedical applications have not been investigated using modern molecular techniques. The proposed work will address this significant knowledge gap through three specific aims. First, a comprehensive black widow venom gland transcriptome will be assembled using next-generation sequencing methods to characterize the genes expressed in venom gland tissue. Second, the set of transcripts involved in venom function will be identified from the transcriptome. This will involve the proteomic technique of peptide fingerprinting to determine the proteins in venom samples using mass spectrometry and bioinformatic screens of a transcriptome database. The next-generation sequencing application of Tag-Profiling will also be used to identify genes that exhibit venom-specific patterns of expression. The third specific aim will examine the functional evolution of black widow venom through cross-species comparisons of venom gene variation that quantify patterns of selection, as well as comparative analyses of gene expression. Phylogenetic analyses of functionally important toxin proteins will also reveal the molecular mechanisms that have contributed to the extreme toxicity of black widow spider venom. When completed, the proposed work will provide an unparalleled view of the molecular diversity, evolution and biomedical utility of black widow venom. The results of this project will also generate significant public genomic resources and provide a foundation for future studies aimed at understanding the molecular mechanisms of toxin functional evolution.

Public Health Relevance

The major goal of this proposal is to determine the molecular composition, functional activities and evolution of black widow spider venom. The project has significant relevance for public health because black widow venom is extremely hazardous to humans, but it is also an essential tool in the study of vertebrate neurophysiology. The proposed work will provide an unprecedented understanding of the molecular basis for the extreme toxicity of black widow spiders and will aid in the development of novel pharmaceuticals and improved antivenoms.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Academic Research Enhancement Awards (AREA) (R15)
Project #
1R15GM097714-01
Application #
8100906
Study Section
Genetic Variation and Evolution Study Section (GVE)
Program Officer
Eckstrand, Irene A
Project Start
2011-04-01
Project End
2015-03-31
Budget Start
2011-04-01
Budget End
2015-03-31
Support Year
1
Fiscal Year
2011
Total Cost
$294,656
Indirect Cost
Name
University of Massachusetts Lowell
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
956072490
City
Lowell
State
MA
Country
United States
Zip Code
01854
Gendreau, Kerry L; Haney, Robert A; Schwager, Evelyn E et al. (2017) House spider genome uncovers evolutionary shifts in the diversity and expression of black widow venom proteins associated with extreme toxicity. BMC Genomics 18:178
Vienneau-Hathaway, Jannelle M; Brassfield, Elizabeth R; Lane, Amanda Kelly et al. (2017) Duplication and concerted evolution of MiSp-encoding genes underlie the material properties of minor ampullate silks of cobweb weaving spiders. BMC Evol Biol 17:78
Haney, Robert A; Clarke, Thomas H; Gadgil, Rujuta et al. (2016) Effects of Gene Duplication, Positive Selection, and Shifts in Gene Expression on the Evolution of the Venom Gland Transcriptome in Widow Spiders. Genome Biol Evol 8:228-42
Clarke, Thomas H; Garb, Jessica E; Hayashi, Cheryl Y et al. (2015) Spider Transcriptomes Identify Ancient Large-Scale Gene Duplication Event Potentially Important in Silk Gland Evolution. Genome Biol Evol 7:1856-70
Clarke, Thomas H; Garb, Jessica E; Hayashi, Cheryl Y et al. (2014) Multi-tissue transcriptomics of the black widow spider reveals expansions, co-options, and functional processes of the silk gland gene toolkit. BMC Genomics 15:365
Bhere, Kanaka Varun; Haney, Robert A; Ayoub, Nadia A et al. (2014) Gene structure, regulatory control, and evolution of black widow venom latrotoxins. FEBS Lett 588:3891-7
Garb, Jessica E (2014) Extraction of venom and venom gland microdissections from spiders for proteomic and transcriptomic analyses. J Vis Exp :e51618
Haney, Robert A; Ayoub, Nadia A; Clarke, Thomas H et al. (2014) Dramatic expansion of the black widow toxin arsenal uncovered by multi-tissue transcriptomics and venom proteomics. BMC Genomics 15:366
McCowan, Caryn; Garb, Jessica E (2014) Recruitment and diversification of an ecdysozoan family of neuropeptide hormones for black widow spider venom expression. Gene 536:366-75
Garb, Jessica E; Hayashi, Cheryl Y (2013) Molecular evolution of ?-latrotoxin, the exceptionally potent vertebrate neurotoxin in black widow spider venom. Mol Biol Evol 30:999-1014