The laccase multi-gene family in Coprinopsis cinerea has seventeen different members that divide into two distinct subfamilies
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Seventeen non-allelic laccase genes and one gene footprint are present in the genome of Coprinopsis cinerea. Two gene subfamilies were defined by intron positions and similarity of deduced gene products, one with 15 members (lcc1–lcc15) and one with 2 members (lcc16, lcc17). The first subfamily divides in the phylogenetic tree of deduced proteins into smaller clusters that probably reflect recent gene duplication events. Different laccase genes diverged from each other both by frequent synonymous and non-synonymous codon changes. Mainly synonymous codon changes accumulate in alleles, with up to 12% total codon differences between given pairs of alleles. Overexpression of the 17 laccase genes under the control of a constitutive promoter identified nine active enzymes from subfamily 1. All of these showed laccase activities with DMP (2,6-dimethoxy phenol) as substrate but only eight of them also with ABTS [2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)]. Lcc16 and Lcc17 share certain sequence features with ferroxidases but enzyme assays failed to show such activity. Lcc15 is expected to be non-functional in laccase activity due to an internal deletion of about 150 amino acids. Transcripts were obtained from all genes but splice junctions for three genes were not congruent with translation into a functional protein.
KeywordsLaccase subfamilies Multi copper oxidases Gene duplication Genome evolution Synonymous and non-synonymous codon changes Splicing
We very much appreciate Tim James and Jason Thacker for valuable discussions and critical comments on this manuscript and Allen Gathman for including the deduced gene sequences into the Coprinus cinereus genome annotation database. We thank the Broad Institute for releasing to the public the sequence of C. cinerea strain Okayama 7 (http://www.genome.wi.mit.edu/annotation/fungi/coprinus_cinereus/index.html), the Göttingen Genomics Laboratory (Göttingen, Germany) for providing their sequencing services. Our laboratory is supported by the Deutsche Bundesstiftung Umwelt (DBU).
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