A transcribed polyketide synthase gene from Xanthoria elegans.
We characterize the transcript of a polyketide synthase gene (PKS) from the cultured mycobiont of Xanthoria elegans (XePKS1) using SMART-rapid amplification of cDNA ends (RACE) cDNA synthesis. Sequence analysis of the cloned cDNA reveals an open reading frame of 2144 amino acid residues. It contains features of a non-reducing fungal type I PKS with an N-terminal starter unit: acyl carrier protein (ACP) transacetylase domain, ketosynthase, acyltransferase, two acyl carrier protein domains, and a thioesterase domain. XePKS1 was the only paralogue detected in the cDNA and the genomic DNA of the cultured X. elegans mycobiont by using a degenerate PCR approach targeted at the conserved regions of non-reducing type I PKS genes. The hypothetical protein is phylogenetically related to genes that are basal to a clade of dihydroxynaphthalene synthases (non-reducing clade II) and anthraquinone type synthases of non-lichenized fungi (non-reducing clade I). According to hplc and tlc analyses, the cultured mycobiont exclusively produced anthraquinones and its precursors. Therefore, we discuss whether the characterized paralogue is involved in anthraquinone production, which raises the possibility of a paraphyletic origin of lichen anthraquinone biosynthesis. The cDNA of XePKS1 was the first full-length coding sequence of a lichen PKS to be published. This proves SMART RACE to be a suitable tool for obtaining full-length coding sequences of genes from environmental samples and organisms, which are hardly amenable to standard molecular approaches or genomic sequencing.
Brunauer
Georg
G
Department of Organismic Biology, University of Salzburg, Hellbrunnerstr. 34, 5020 Salzburg, Austria. georg.brunauer@sbg.ac.at
Muggia
Lucia
L
Stocker-Wörgötter
Elfie
E
Grube
Martin
M
D000595:Amino Acid Sequence; D000880:Anthraquinones / Q000378:metabolism; D001203:Ascomycota / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D003470:Culture Media; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism*; D008011:Lichens / Q000382:microbiology*; D008969:Molecular Sequence Data; D010802:Phylogeny; D048630:Polyketide Synthases / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012331:RNA, Fungal / Q000032:analysis / Q000302:isolation & purification; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D000880:Anthraquinones; D003470:Culture Media; D017931:DNA Primers; D018076:DNA, Complementary; D012331:RNA, Fungal; D048630:Polyketide Synthases