title: Evolution of the relaxin-like peptide family.
abstract: BACKGROUND
  The relaxin-like peptide family belongs in the insulin superfamily and consists of 7 peptides of high structural but low sequence similarity; relaxin-1, 2 and 3, and the insulin-like (INSL) peptides, INSL3, INSL4, INSL5 and INSL6. The functions of relaxin-3, INSL4, INSL5, INSL6 remain uncharacterised. The evolution of this family has been contentious; high sequence variability is seen between closely related species, while distantly related species show high similarity; an invertebrate relaxin sequence has been reported, while a relaxin gene has not been found in the avian and ruminant lineages.
  
  
  RESULTS
  Sequence similarity searches of genomic and EST data identified homologs of relaxin-like peptides in mammals, and non-mammalian vertebrates such as fish. Phylogenetic analysis was used to resolve the evolution of the family. Searches were unable to identify an invertebrate relaxin-like peptide. The published relaxin cDNA sequence in the tunicate, Ciona intestinalis was not present in the completed C. intestinalis genome. The newly discovered relaxin-3 is likely to be the ancestral relaxin. Multiple relaxin-3-like sequences are present in fugu fish (Takifugu rubripes) and zebrafish (Danio rerio), but these appear to be specific to the fish lineage. Possible relaxin-1 and INSL5 homologs were also identified in fish and frog species, placing their emergence prior to mammalia, earlier than previously believed. Furthermore, estimates of synonymous and nonsynonymous substitution rates (dN/dS) suggest that the emergence of relaxin-1, INSL4 and INSL6 during mammalia was driven by positive Darwinian selection, hence these peptides are likely to have novel and in the case of relaxin-1, which is still under positive selection in humans and the great apes, possibly still evolving functions. In contrast, relaxin-3 is constrained by strong purifying selection, demonstrating it must have a highly conserved function, supporting its hypothesized important neuropeptide role.
  
  
  CONCLUSIONS
  We present a phylogeny describing the evolutionary history of the relaxin-like peptide family and show that positive selection has driven the evolution of the most recent members of the family.
authors.affiliation: Howard Florey Institute of Experimental Physiology and Medicine, University of Melbourne, Australia. t.wilkinson@hfi.unimelb.edu.au
authors.forename: Tracey N
authors.initials: TN
authors.lastname: Wilkinson
authors.forename: Terence P
authors.initials: TP
authors.lastname: Speed
authors.forename: Geoffrey W
authors.initials: GW
authors.lastname: Tregear
authors.forename: Ross A D
authors.initials: RA
authors.lastname: Bathgate
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D004285:Dogs; D019143:Evolution, Molecular; D006801:Humans; D007328:Insulin / Q000145:classification* / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D010802:Phylogeny; D051381:Rats; D012065:Relaxin / Q000145:classification* / Q000235:genetics*; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein
chemical_list: D007328:Insulin; D012065:Relaxin