Avtor/Urednik | Lanišnik-Rižner, Tea; Wheeler, Michael H | |
Naslov | Melanin biosynthesis in the fungus Curvularia lunata (teleomorph: Cochliobolus lunatus) | |
Tip | članek | |
Vir | Can J Microbiol | |
Vol. in št. | Letnik 49 | |
Leto izdaje | 2003 | |
Obseg | str. 110-9 | |
Jezik | eng | |
Abstrakt | Curvularia lunata (teleomorph: Cochliobolus lunatus) is a known plant and human pathogen. Tricyclazole, a specific inhibitor of pentaketide melanin biosynthesis, blocked the biosynthesis of melanin in Curvularia lunata and caused the accumulation of the melanin metabolites flaviolin and 2-hydroxyjuglone. This showed that melanin in Curvularia lunata is produced by a pentaketide pathway from 1,8-dihydroxynaphthalene. The 1,3,8trihydroxynaphthalene reductase (3HNR) gene, associated with the melanin pathway of Curvularia lunata, was identified and characterized. An alignment of 3HNR sequences enabled the design of primers covering conserved regions. A PCR-amplified fragment of Curvularia lunata genomic DNA was used for screening the cDNA library. Three independent cDNA clones revealed an 801-bp open reading frame encoding a 267 amino acid protein. The protein was expressed in Escherichia coli and purified to homogeneity. The predicted amino acid sequence of the 28.6-kDa protein demonstrated homology to other fungal 3HNR and other members of the short-chain dehydrogenase super family. Northern analyses revealed that 3HNR from Curvularia lunata is expressed synchronously with melanization after 3 days of Curvularia lunata growth in malt extract medium. No 3HNR reductase gene expression nor melanization was observed when Curvularia lunata was grown in yeast nitrogen base medium. | |
Deskriptorji | ASCOMYCETES MELANINS NAPHTHOLS OXIDOREDUCTASES DNA, FUNGAL POLYMERASE CHAIN REACTION GENE LIBRARY AMINO ACID SEQUENCE DNA, COMPLEMENTARY GENOME, FUNGAL CHROMATOGRAPHY, THIN LAYER CHROMATOGRAPHY, HIGH PRESSURE LIQUID BLOTTING, NORTHERN SPECTROPHOTOMETRY |