Inventing an arsenal: adaptive evolution and neofunctionalization of snake venom phospholipase A2 genes

BackgroundGene duplication followed by functional divergence has long been hypothesized to be the main source of molecular novelty. Convincing examples of neofunctionalization, however, remain rare. Snake venom phospholipase A2 genes are members of large multigene families with many diverse functions, thus they are excellent models to study the emergence of novel functions after gene duplications.ResultsHere, I show that positive Darwinian selection and neofunctionalization is common in snake venom phospholipase A2 genes. The pattern of gene duplication and positive selection indicates that adaptive molecular evolution occurs immediately after duplication events as novel functions emerge and continues as gene families diversify and are refined. Surprisingly, adaptive evolution of group-I phospholipases in elapids is also associated with speciation events, suggesting adaptation of the phospholipase arsenal to novel prey species after niche shifts. Mapping the location of sites under positive selection onto the crystal structure of phospholipase A2 identified regions evolving under diversifying selection are located on the molecular surface and are likely protein-protein interactions sites essential for toxin functions.ConclusionThese data show that increases in genomic complexity (through gene duplications) can lead to phenotypic complexity (venom composition) and that positive Darwinian selection is a common evolutionary force in snake venoms. Finally, regions identified under selection on the surface of phospholipase A2 enzymes are potential candidate sites for structure based antivenin design.

[1]  Y. Belyi Phospholipases. , 1991, Methods in enzymology.

[2]  N. Guex,et al.  SWISS‐MODEL and the Swiss‐Pdb Viewer: An environment for comparative protein modeling , 1997, Electrophoresis.

[3]  R. Kini Excitement ahead: structure, function and mechanism of snake venom phospholipase A2 enzymes. , 2003, Toxicon : official journal of the International Society on Toxinology.

[4]  EDWIN C. Webb The Enzymes , 1961, Nature.

[5]  N. Goldman,et al.  A codon-based model of nucleotide substitution for protein-coding DNA sequences. , 1994, Molecular biology and evolution.

[6]  M. Tu,et al.  Food Habits of the Sea Snake, Laticauda semifasciata , 2005 .

[7]  A F Bennett,et al.  Genetic architecture of thermal adaptation in Escherichia coli. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[8]  R. Kini,et al.  The role of enzymatic activity in inhibition of the extrinsic tenase complex by phospholipase A2 isoenzymes from Naja nigricollis venom. , 1995, Toxicon : official journal of the International Society on Toxinology.

[9]  Ziheng Yang,et al.  Statistical methods for detecting molecular adaptation , 2000, Trends in Ecology & Evolution.

[10]  R. Kini Venom phospholipase A[2] enzymes : structure, function, and mechanism , 1997 .

[11]  R. Kini,et al.  A model to explain the pharmacological effects of snake venom phospholipases A2. , 1989, Toxicon : official journal of the International Society on Toxinology.

[12]  Z. Yang,et al.  Likelihood ratio tests for detecting positive selection and application to primate lysozyme evolution. , 1998, Molecular biology and evolution.

[13]  D. Kordis,et al.  Positive Darwinian selection in Vipera palaestinae phospholipase A2 genes is unexpectedly limited to the third exon. , 1998, Biochemical and biophysical research communications.

[14]  A. Soares,et al.  Chemical modifications of phospholipases A2 from snake venoms: effects on catalytic and pharmacological properties. , 2003, Toxicon : official journal of the International Society on Toxinology.

[15]  Ziheng Yang,et al.  PAML: a program package for phylogenetic analysis by maximum likelihood , 1997, Comput. Appl. Biosci..

[16]  John P. Huelsenbeck,et al.  MrBayes 3: Bayesian phylogenetic inference under mixed models , 2003, Bioinform..

[17]  H. Verheij,et al.  The role of aspartic acid-49 in the active site of phospholipase A2. A site-specific mutagenesis study of porcine pancreatic phospholipase A2 and the rationale of the enzymatic activity of [lysine49]phospholipase A2 from Agkistrodon piscivorus piscivorus' venom. , 1988, European journal of biochemistry.

[18]  Y. Kariya,et al.  Nucleotide sequence of phospholipase A(2) gene expressed in snake pancreas reveals the molecular evolution of toxic phospholipase A(2) genes. , 2002, Gene.

[19]  N. Goldman,et al.  Codon-substitution models for heterogeneous selection pressure at amino acid sites. , 2000, Genetics.

[20]  R. Shine,et al.  Relationships between sexual dimorphism and niche partitioning within a clade of sea-snakes (Laticaudinae) , 2002, Oecologia.

[21]  Dr. Susumu Ohno Evolution by Gene Duplication , 1970, Springer Berlin Heidelberg.

[22]  Yiu Man Chan,et al.  Accelerated Evolution and Molecular Surface of Venom Phospholipase A2 Enzymes , 1999, Journal of Molecular Evolution.

[23]  A. Knight,et al.  Inferring species trees from gene trees: a phylogenetic analysis of the Elapidae (Serpentes) based on the amino acid sequences of venom proteins. , 1997, Molecular phylogenetics and evolution.

[24]  Miss A.O. Penney (b) , 1974, The New Yale Book of Quotations.

[25]  Y. Chuman,et al.  Regional and accelerated molecular evolution in group I snake venom gland phospholipase A2 isozymes. , 2000, Toxicon : official journal of the International Society on Toxinology.

[26]  R. Shine Habitats, diets, and sympatry in snakes: a study from Australia , 1977 .

[27]  R. Ward,et al.  Mapping structural determinants of biological activities in snake venom phospholipases A2 by sequence analysis and site directed mutagenesis. , 2003, Toxicon : official journal of the International Society on Toxinology.

[28]  J. Gené,et al.  The effect of myotoxins isolated from Bothrops snake venoms on multilamellar liposomes: relationship to phospholipase A2, anticoagulant and myotoxic activities. , 1991, Biochimica et biophysica acta.

[29]  M. Hattori,et al.  Accelerated evolution in the protein-coding regions is universal in crotalinae snake venom gland phospholipase A2 isozyme genes. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[30]  Jianzhi Zhang,et al.  Adaptive evolution of a duplicated pancreatic ribonuclease gene in a leaf-eating monkey , 2002, Nature Genetics.

[31]  A. Desideri,et al.  Calcium ion independent membrane leakage induced by phospholipase-like myotoxins. , 1992, Biochemistry.

[32]  Vladimir Brusic,et al.  Molecular Evolution and Phylogeny of Elapid Snake Venom Three-Finger Toxins , 2003, Journal of Molecular Evolution.

[33]  R. Heinrikson,et al.  The lysine-49 phospholipase A2 from the venom of Agkistrodon piscivorus piscivorus. Relation of structure and function to other phospholipases A2. , 1986, The Journal of biological chemistry.

[34]  Isd Habitats, diets and sympatry in snakes: a study from Australia , 1978 .

[35]  J. Thompson,et al.  CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. , 1994, Nucleic acids research.