Full-Length Venom Protein cDNA Sequences from Venom-Derived mRNA: Exploring Compositional Variation and Adaptive Multigene Evolution.
Envenomation of humans by snakes is a complex and continuously evolving medical emergency, and treatment is made that much more difficult by the diverse biochemical composition of many venoms. Venomous snakes and their venoms also provide models for the study of molecular evolutionary processes leading to adaptation and genotype-phenotype relationships. To compare venom complexity and protein sequences, venom gland transcriptomes are assembled, which usually requires the sacrifice of snakes for tissue. However, toxin transcripts are also present in venoms, offering the possibility of obtaining cDNA sequences directly from venom. This study provides evidence that unknown full-length venom protein transcripts can be obtained from the venoms of multiple species from all major venomous snake families. These unknown venom protein cDNAs are obtained by the use of primers designed from conserved signal peptide sequences within each venom protein superfamily. This technique was used to assemble a partial venom gland transcriptome for the Middle American Rattlesnake (Crotalus simus tzabcan) by amplifying sequences for phospholipases A2, serine proteases, C-lectins, and metalloproteinases from within venom. Phospholipase A2 sequences were also recovered from the venoms of several rattlesnakes and an elapid snake (Pseudechis porphyriacus), and three-finger toxin sequences were recovered from multiple rear-fanged snake species, demonstrating that the three major clades of advanced snakes (Elapidae, Viperidae, Colubridae) have stable mRNA present in their venoms. These cDNA sequences from venom were then used to explore potential activities derived from protein sequence similarities and evolutionary histories within these large multigene superfamilies. Venom-derived sequences can also be used to aid in characterizing venoms that lack proteomic profiles and identify sequence characteristics indicating specific envenomation profiles. This approach, requiring only venom, provides access to cDNA sequences in the absence of living specimens, even from commercial venom sources, to evaluate important regional differences in venom composition and to study snake venom protein evolution.
School of Biological Sciences, University of Northern Colorado, Greeley, Colorado, United States of America.
Cassandra M
CM
Modahl
School of Biological Sciences, University of Northern Colorado, Greeley, Colorado, United States of America.
Stephen P
SP
Mackessy
D000222:Adaptation, Physiological; D000595:Amino Acid Sequence; D000818:Animals; D005075:Biological Evolution; D003001:Cloning, Molecular; D017812:Colubridae / Q000502:physiology*; D018076:DNA, Complementary / Q000235:genetics*; D004546:Elapid Venoms / Q000737:chemistry / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D014757:Viper Venoms / Q000737:chemistry / Q000235:genetics*; D017819:Viperidae / Q000502:physiology*
D018076:DNA, Complementary; D004546:Elapid Venoms; D012333:RNA, Messenger; D014757:Viper Venoms