title: Opossum carboxylesterases: sequences, phylogeny and evidence for CES gene duplication events predating the marsupial-eutherian common ancestor.
abstract: BACKGROUND
  Carboxylesterases (CES) perform diverse metabolic roles in mammalian organisms in the detoxification of a broad range of drugs and xenobiotics and may also serve in specific roles in lipid, cholesterol, pheromone and lung surfactant metabolism. Five CES families have been reported in mammals with human CES1 and CES2 the most extensively studied. Here we describe the genetics, expression and phylogeny of CES isozymes in the opossum and report on the sequences and locations of CES1, CES2 and CES6 'like' genes within two gene clusters on chromosome one. We also discuss the likely sequence of gene duplication events generating multiple CES genes during vertebrate evolution.
  
  
  RESULTS
  We report a cDNA sequence for an opossum CES and present evidence for CES1 and CES2 like genes expressed in opossum liver and intestine and for distinct gene locations of five opossum CES genes,CES1, CES2.1, CES2.2, CES2.3 and CES6, on chromosome 1. Phylogenetic and sequence alignment studies compared the predicted amino acid sequences for opossum CES with those for human, mouse, chicken, frog, salmon and Drosophila CES gene products. Phylogenetic analyses produced congruent phylogenetic trees depicting a rapid early diversification into at least five distinct CES gene family clusters: CES2, CES1, CES7, CES3, and CES6. Molecular divergence estimates based on a Bayesian relaxed clock approach revealed an origin for the five mammalian CES gene families between 328-378 MYA.
  
  
  CONCLUSIONS
  The deduced amino acid sequence for an opossum cDNA was consistent with its identity as a mammalian CES2 gene product (designated CES2.1). Distinct gene locations for opossum CES1 (1: 446,222,550-446,274,850), three CES2 genes (1: 677,773,395-677,927,030) and a CES6 gene (1: 677,585,520-677,730,419) were observed on chromosome 1. Opossum CES1 and multiple CES2 genes were expressed in liver and intestine. Amino acid sequences for opossum CES1 and three CES2 gene products revealed conserved residues previously reported for human CES1 involved in catalysis, ligand binding, tertiary structure and organelle localization. Phylogenetic studies indicated the gene duplication events which generated ancestral mammalian CES genes predated the common ancestor for marsupial and eutherian mammals, and appear to coincide with the early diversification of tetrapods.
authors.affiliation: Department of Genetics, Southwest Foundation for Biomedical Research, San Antonio, TX, USA. rholmes@sfbrgenetics.org
authors.forename: Roger S
authors.initials: RS
authors.lastname: Holmes
authors.forename: Jeannie
authors.initials: J
authors.lastname: Chan
authors.forename: Laura A
authors.initials: LA
authors.lastname: Cox
authors.forename: William J
authors.initials: WJ
authors.lastname: Murphy
authors.forename: John L
authors.initials: JL
authors.lastname: VandeBerg
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000235:genetics*; D002874:Chromosome Mapping; D018076:DNA, Complementary; D019143:Evolution, Molecular; D020440:Gene Duplication; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007422:Intestines / Q000201:enzymology; D007527:Isoenzymes / Q000235:genetics; D008099:Liver / Q000201:enzymology; D009893:Opossums / Q000235:genetics* / Q000378:metabolism; D009928:Organ Specificity; D010802:Phylogeny; D017433:Protein Structure, Secondary; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D013045:Species Specificity; D013379:Substrate Specificity
chemical_list: D018076:DNA, Complementary; D007527:Isoenzymes; D002265:Carboxylic Ester Hydrolases