title: Tandem cytochrome P450 monooxygenase genes and splice variants in the white rot fungus Phanerochaete chrysosporium: cloning, sequence analysis, and regulation of differential expression.
abstract: Phanerochaete chrysosporium, the model white rot fungus, uses P450 monooxygenation for oxidation of xenobiotic chemicals. To characterize its P450 systems, we have isolated two tandemly linked full-length P450 monooxygenase genes, pc-1 (2510bp) and pc-2 (2503bp). cDNA sequence and splice site analysis showed multiple small putative introns (35-82bp), 17 in pc-1 and 16 in pc-2. The deduced P450 proteins PC-1 (531 aa, MW 60,070) and PC-2 (538 aa, MW 61,765) showed 55% aa identity with each other and low relatedness (25-36%) with P450 gene families CYP52, CYP86, CYP4, and CYP3 decreasing in that order. Alternate splicing was identified for pc-1 transcription. Real-time quantitative RT-PCR analysis on 4-day cultures showed an elevated level of expression ( approximately 8-fold) of pc-1 in nutrient-limited (ligninolytic) as compared to nutrient-rich (non-ligninolytic) conditions. Interestingly, the pc-1 gene is expressible in both primary and secondary phases of growth with peak expression occurring with secondary metabolism.
authors.affiliation: Molecular Toxicology Division, Department of Environmental Health, College of Medicine, University of Cincinnati, Cincinnati, OH 45267-0056, USA. Jagjit.Yadav@uc.edu
authors.forename: Jagjit S
authors.initials: JS
authors.lastname: Yadav
authors.forename: Matthew B
authors.initials: MB
authors.lastname: Soellner
authors.forename: John C
authors.initials: JC
authors.lastname: Loper
authors.forename: Prashant K
authors.initials: PK
authors.lastname: Mishra
mesh_terms: D017398:Alternative Splicing / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D001487:Basidiomycota / Q000235:genetics; D001673:Biodegradation, Environmental; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics*; D004274:DNA, Recombinant; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D008969:Molecular Sequence Data; D020075:Phanerochaete / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid
chemical_list: D018076:DNA, Complementary; D004274:DNA, Recombinant; D003577:Cytochrome P-450 Enzyme System