title: Computational analysis and functional expression of ancestral copepod luciferase.
abstract: We recently reported the cDNA sequences of 11 copepod luciferases from the superfamily Augaptiloidea in the order Calanoida. They were classified into two groups, Metridinidae and Heterorhabdidae/Lucicutiidae families, by phylogenetic analyses. To elucidate the evolutionary processes, we have now further isolated 12 copepod luciferases from Augaptiloidea species (Metridia asymmetrica, Metridia curticauda, Pleuromamma scutullata, Pleuromamma xiphias, Lucicutia ovaliformis and Heterorhabdus tanneri). Codon-based synonymous/nonsynonymous tests of positive selection for 25 identified copepod luciferases suggested that positive Darwinian selection operated in the evolution of Heterorhabdidae luciferases, whereas two types of Metridinidae luciferases had diversified via neutral mechanism. By in silico analysis of the decoded amino acid sequences of 25 copepod luciferases, we inferred two protein sequences as ancestral copepod luciferases. They were expressed in HEK293 cells where they exhibited notable luciferase activity both in intracellular lysates and cultured media, indicating that the luciferase activity was established before evolutionary diversification of these copepod species.
authors.affiliation: Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan. pc_tert0617@yahoo.co.jp
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Takenaka
authors.forename: Akiko
authors.initials: A
authors.lastname: Noda-Ogura
authors.forename: Tadashi
authors.initials: T
authors.lastname: Imanishi
authors.forename: Atsushi
authors.initials: A
authors.lastname: Yamaguchi
authors.forename: Takashi
authors.initials: T
authors.lastname: Gojobori
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Shigeri
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D060829:Arthropod Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D019295:Computational Biology; D033342:Copepoda / Q000201:enzymology*; D019143:Evolution, Molecular; D015870:Gene Expression; D049810:Genetic Speciation; D057809:HEK293 Cells; D006801:Humans; D008156:Luciferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid
chemical_list: D060829:Arthropod Proteins; D008156:Luciferases
keywords: ASR; Ancestral gene; Bioluminescence; Coe; Coelenterazine; Copepod; GLuc; Gaussia luciferase; Luciferase; Luciferin; ML; MLuc; Metridia longa luciferase; ancestral sequence reconstruction; coelenterazine; maximum likelihood