title: Characterization of the Aspergillus ochraceoroseus aflatoxin/sterigmatocystin biosynthetic gene cluster.
abstract: Production of carcinogenic aflatoxins has been reported from members of Aspergillus section Flavi, Aspergillus section Nidulantes and a newly proposed Aspergillus section Ochraceorosei that consists of Aspergillus ochraceoroseus and A. rambellii. Unlike members of section Flavi, A. ochraceoroseus and A. rambellii have been shown to accumulate both aflatoxin (AF) and the aflatoxin precursor sterigmatocystin (ST). Alhough morphologically distinct from A. nidulans, molecular characterization of A. ochraceoroseus AF/ST genes and physiological characteristics of AF/ST production indicated that A. ochraceoroseus is more closely related to A. nidulans than to A. flavus. Knowing that the A. nidulans ST gene cluster is organized differently from the A. flavus AF gene cluster, we determined the genetic organization of the AF/ST biosynthetic cluster in A. ochraceoroseus. Sequencing of overlapping lambda clones and genomic PCR fragments obtained by gene-walking techniques demonstrated that the A. ochraceoroseus AF/ST gene cluster is organized much like the A. nidulans ST gene cluster except that the region from aflN to aflW is located directly upstream of aflC and in reverse orientation such that aflW represents the distal end and aflY the proximal end of the cluster. The A. ochraceoroseus cluster genes demonstrated 62-76% nucleotide identity to their A. nidulans ST cluster gene homologs. Transformation of an A. nidulans aflR mutant with the A. ochraceoroseus aflR restored ST production in A. nidulans transformants. PCR amplification of A. rambellii genomic DNA demonstrated that the AF/ST gene cluster is organized in the same manner as that of A. ochraceoroseus.
authors.affiliation: USDA, ARS, Southern Regional Research Center, 1100 Robert E. Lee Boulevard, New Orleans, Louisiana 70124, USA. Jeff.Cary@ars.usda.gov
authors.forename: J W
authors.initials: JW
authors.lastname: Cary
authors.forename: K C
authors.initials: KC
authors.lastname: Ehrlich
authors.forename: S B
authors.initials: SB
authors.lastname: Beltz
authors.forename: P
authors.initials: P
authors.lastname: Harris-Coward
authors.forename: M A
authors.initials: MA
authors.lastname: Klich
mesh_terms: D000348:Aflatoxins / Q000096:biosynthesis / Q000235:genetics*; D001231:Aspergillus flavus / Q000235:genetics / Q000378:metabolism; D001233:Aspergillus nidulans / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D001235:Aspergillus ochraceus / Q000235:genetics* / Q000378:metabolism; D015152:Blotting, Northern; D003517:Cyclopentanes / Q000494:pharmacology; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D005656:Fungal Proteins / Q000096:biosynthesis / Q000235:genetics; D014644:Genetic Variation; D005810:Multigene Family; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D013241:Sterigmatocystin / Q000096:biosynthesis*; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics; D015533:Transcriptional Activation / Q000187:drug effects
chemical_list: C113837:AFLR protein, Aspergillus; D000348:Aflatoxins; D003517:Cyclopentanes; D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D014157:Transcription Factors; D013241:Sterigmatocystin; C007201:cyclopentanone