title: Large gene family expansion and variable selective pressures for cathepsin B in aphids.
abstract: Aphids exclusively feed on plant phloem sap that contains much sugar and some nonessential amino acids but is poor in lipids and proteins. Conventionally, it has been believed that aphids substantially have no intestinal digestion of proteins. However, we here report an unexpected finding that cysteine protease genes of the family cathepsin B are massively amplified in the lineage of aphids and that many of the protease genes exhibit gut-specific overexpression. By making use of expressed sequence tag data, sequenced cDNAs, and genomic trace sequences of the pea aphid Acyrthosiphon pisum, we identified a total of 28 cathepsin B-like gene copies in the genome of A. pisum. Phylogenetic analyses of all the cathepsin B genes in aphids revealed that genic expansion has continuously proceeded with basal, intermediary, and recent duplications. Estimation of molecular evolutionary rates indicated that major alterations of the rates often occurred after duplications. For example, a gene copy ("348") was shown to be slow evolving and close to genes of other insects like Drosophila melanogaster, whereas the other gene copies appeared to have evolved faster with higher ratios of nonsynonymous to synonymous substitutions. We identified a number of gene copies (16 in A. pisum) that contained a replacement at the site required for catalytic activity of the protease. Among these, 2 copies were pseudogenes, whereas the remaining copies were structurally intact and possibly acquired new functions. For example, a cluster of such gene copies ("1674") has been subjected to positive selection. Quantitative reverse transcriptase-polymerase chain reaction analyses revealed that the more conserved gene copy ("348") showed a constitutive expression, whereas 5 other forms ("84," "16," "16D," "1874," and "2744") were preferentially expressed in the gut of A. pisum. Putative biological roles of the diversified cathepsin B-like gene copies in aphids are discussed in relation to their nutritional physiology specialized for plant sap feeding lifestyle.
authors.affiliation: Institut National de la Recherche Agronomique, Unité Mixte de Recherche 1099 BIO3P, Le Rheu, France. claude.rispe@rennes.inra.fr
authors.forename: Claude
authors.initials: C
authors.lastname: Rispe
authors.forename: Mayako
authors.initials: M
authors.lastname: Kutsukake
authors.forename: Vincent
authors.initials: V
authors.lastname: Doublet
authors.forename: Sylvie
authors.initials: S
authors.lastname: Hudaverdian
authors.forename: Fabrice
authors.initials: F
authors.lastname: Legeai
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Simon
authors.forename: Denis
authors.initials: D
authors.lastname: Tagu
authors.forename: Takema
authors.initials: T
authors.lastname: Fukatsu
mesh_terms: D000818:Animals; D001042:Aphids / Q000235:genetics* / Q000378:metabolism; D056649:Cathepsin G; D002403:Cathepsins / Q000235:genetics* / Q000378:metabolism; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism; D005247:Feeding Behavior / Q000502:physiology; D018628:Gene Dosage / Q000502:physiology*; D049750:Genome, Insect / Q000502:physiology*; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D005810:Multigene Family / Q000502:physiology*; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism
chemical_list: D019476:Insect Proteins; D002403:Cathepsins; D012697:Serine Endopeptidases; D056649:Cathepsin G