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Linking the transcriptome and proteome to characterize the venom of the eastern diamondback rattlesnake (Crotalus adamanteus).
J Proteomics. 2014 Jan 16; 96:145-58.JP

Abstract

Understanding the molecular basis of the phenotype is key to understanding adaptation, and the relationship between genes and specific traits is represented by the genotype-phenotype map. The specialization of the venom-gland towards toxin production enables the use of transcriptomics to identify a large number of loci that contribute to a complex phenotype (i.e., venom), while proteomic techniques allow verification of the secretion of the proteins produced by these loci, creating a genotype-phenotype map. We used the extensive database of mRNA transcripts generated by the venom-gland transcriptome of Crotalus adamanteus along with proteomic techniques to complete the genotype-phenotype map for the C. adamanteus venom system. Nanospray LC/MS(E) analysis of a whole venom sample identified evidence for 52 of the 78 unique putative toxin transcript clusters, including 44 of the 50 most highly expressed transcripts. Tandem mass spectrometry and SDS-PAGE of reversed-phase high-performance liquid chromatography fractions identified 40 toxins which clustered into 20 groups and represented 10 toxin families, creating a genotype-phenotype map. By using the transcriptome to understand the proteome we were able to achieve locus-specific resolution and provide a detailed characterization of the C. adamanteus venom system.

BIOLOGICAL SIGNIFICANCE

Identifying the mechanisms by which genetic variation presents itself to the sieve of selection at the phenotypic level is key to understanding the molecular basis of adaptation, and the first step in understanding this relationship is to identify the genetic basis of the phenotype through the construction of a genotype-phenotype map. We used the high-throughput venom-gland transcriptomic characterization of the eastern diamondback rattlesnake (C. adamanteus) and proteomic techniques to complete and confirm the genotype-phenotype map, providing a detailed characterization of the C. adamanteus venom system.

Authors+Show Affiliations

Department of Biological Science, Florida State University, Tallahassee, FL, USA.Department of Biological Science, Florida State University, Tallahassee, FL, USA.Department of Biological Science, Florida State University, Tallahassee, FL, USA.Department of Biological Science, Florida State University, Tallahassee, FL, USA.College of Medicine, Florida State University, Tallahassee, FL, USA.Department of Biological Science, Florida State University, Tallahassee, FL, USA. Electronic address: drokyta@bio.fsu.edu.

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

24231107

Citation

Margres, Mark J., et al. "Linking the Transcriptome and Proteome to Characterize the Venom of the Eastern Diamondback Rattlesnake (Crotalus Adamanteus)." Journal of Proteomics, vol. 96, 2014, pp. 145-58.
Margres MJ, McGivern JJ, Wray KP, et al. Linking the transcriptome and proteome to characterize the venom of the eastern diamondback rattlesnake (Crotalus adamanteus). J Proteomics. 2014;96:145-58.
Margres, M. J., McGivern, J. J., Wray, K. P., Seavy, M., Calvin, K., & Rokyta, D. R. (2014). Linking the transcriptome and proteome to characterize the venom of the eastern diamondback rattlesnake (Crotalus adamanteus). Journal of Proteomics, 96, 145-58. https://doi.org/10.1016/j.jprot.2013.11.001
Margres MJ, et al. Linking the Transcriptome and Proteome to Characterize the Venom of the Eastern Diamondback Rattlesnake (Crotalus Adamanteus). J Proteomics. 2014 Jan 16;96:145-58. PubMed PMID: 24231107.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Linking the transcriptome and proteome to characterize the venom of the eastern diamondback rattlesnake (Crotalus adamanteus). AU - Margres,Mark J, AU - McGivern,James J, AU - Wray,Kenneth P, AU - Seavy,Margaret, AU - Calvin,Kate, AU - Rokyta,Darin R, Y1 - 2013/11/12/ PY - 2013/10/01/received PY - 2013/10/25/revised PY - 2013/11/01/accepted PY - 2013/11/16/entrez PY - 2013/11/16/pubmed PY - 2014/9/23/medline KW - Crotalus adamanteus KW - Genotype–phenotype map KW - Liquid chromatography KW - Mass spectrometry KW - Snake venomics KW - Transcriptomics SP - 145 EP - 58 JF - Journal of proteomics JO - J Proteomics VL - 96 N2 - UNLABELLED: Understanding the molecular basis of the phenotype is key to understanding adaptation, and the relationship between genes and specific traits is represented by the genotype-phenotype map. The specialization of the venom-gland towards toxin production enables the use of transcriptomics to identify a large number of loci that contribute to a complex phenotype (i.e., venom), while proteomic techniques allow verification of the secretion of the proteins produced by these loci, creating a genotype-phenotype map. We used the extensive database of mRNA transcripts generated by the venom-gland transcriptome of Crotalus adamanteus along with proteomic techniques to complete the genotype-phenotype map for the C. adamanteus venom system. Nanospray LC/MS(E) analysis of a whole venom sample identified evidence for 52 of the 78 unique putative toxin transcript clusters, including 44 of the 50 most highly expressed transcripts. Tandem mass spectrometry and SDS-PAGE of reversed-phase high-performance liquid chromatography fractions identified 40 toxins which clustered into 20 groups and represented 10 toxin families, creating a genotype-phenotype map. By using the transcriptome to understand the proteome we were able to achieve locus-specific resolution and provide a detailed characterization of the C. adamanteus venom system. BIOLOGICAL SIGNIFICANCE: Identifying the mechanisms by which genetic variation presents itself to the sieve of selection at the phenotypic level is key to understanding the molecular basis of adaptation, and the first step in understanding this relationship is to identify the genetic basis of the phenotype through the construction of a genotype-phenotype map. We used the high-throughput venom-gland transcriptomic characterization of the eastern diamondback rattlesnake (C. adamanteus) and proteomic techniques to complete and confirm the genotype-phenotype map, providing a detailed characterization of the C. adamanteus venom system. SN - 1876-7737 UR - https://www.unboundmedicine.com/medline/citation/24231107/Linking_the_transcriptome_and_proteome_to_characterize_the_venom_of_the_eastern_diamondback_rattlesnake__Crotalus_adamanteus__ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1874-3919(13)00555-1 DB - PRIME DP - Unbound Medicine ER -