The tightly regulated promoter of the xanA gene of Aspergillus nidulans is included in a helitron

In Aspergillus nidulans the xanA gene codes for a xanthine α‐ketoglutarate‐dependent dioxygenase, an enzyme only present in the fungal kingdom. The 5′ region of this gene, including its putative promoter and the first 54 codons of the open reading frame, together with the first intron is duplicated in the genome. This duplication corresponds to a helitron, a eukaryotic element proposed to transpose replicatively by the rolling circle mechanism. We show that the regulation of xanA conforms to that of other genes of the purine degradation pathway, necessitating the specific UaY transcription factor and the AreA GATA factor. The promoter of the duplicated region is active ectopically and the difficulty in detecting an mRNA from the duplicated region is at least partially due to nonsense‐mediated decay. Comparative genomic data are only consistent with the hypothesis that the 5′ region of xanA pre‐existed the helitron insertion, and that a ‘secondary helitron’ was generated from an insertion 5′ to it and a pre‐existing 3′ consensus sequence within the open reading frame. It is possible to propose a role of helitrons in promoter shuffling and thus in recruiting new genes into specific regulatory circuits.

[1]  G. Marzluf Regulation of sulfur and nitrogen metabolism in filamentous fungi. , 1993, Annual review of microbiology.

[2]  W. Timberlake,et al.  Sequence and molecular structure of the Aspergillus nidulans yA (laccase I) gene. , 1990, Nucleic acids research.

[3]  M. Hynes Studies on the role of the areA gene in the regulation of nitrogen catabolism in Aspergillus nidulans. , 1975, Australian journal of biological sciences.

[4]  B. Cohen Regulation of intracellular and extracellular neutral and alkaline proteases in Aspergillus nidulans. , 1973, Journal of general microbiology.

[5]  M. Rochet,et al.  Convergent evolution of hydroxylation mechanisms in the fungal kingdom: molybdenum cofactor‐independent hydroxylation of xanthine via α‐ketoglutarate‐dependent dioxygenases , 2005, Molecular microbiology.

[6]  A. Glatigny,et al.  Cloning and Molecular Characterization of hxA, the Gene Coding for the Xanthine Dehydrogenase (Purine Hydroxylase I) of Aspergillus nidulans(*) , 1995, The Journal of Biological Chemistry.

[7]  C. Scazzocchio,et al.  Sequence and regulation of the uapA gene encoding a uric acid-xanthine permease in the fungus Aspergillus nidulans. , 1993, The Journal of biological chemistry.

[8]  H. Arst,et al.  Nonsense-Mediated mRNA Decay Mutation in Aspergillus nidulans , 2006, Eukaryotic Cell.

[9]  C. Scazzocchio,et al.  The AzgA Purine Transporter of Aspergillus nidulans , 2004, Journal of Biological Chemistry.

[10]  C. Scazzocchio,et al.  The GATA factor AreA is essential for chromatin remodelling in a eukaryotic bidirectional promoter , 1999, The EMBO journal.

[11]  C. Scazzocchio,et al.  The genetic control of molybdoflavoproteins in Aspergillus nidulans. A xanthine dehydrogenase I half-molecule in cnx- mutant strains of Aspergillus nidulans. , 1977, European journal of biochemistry.

[12]  C. Scazzocchio,et al.  Molecular cloning of the uaY regulatory gene of Aspergillus nidulans reveals a favoured region for DNA insertions , 1991, Molecular and General Genetics MGG.

[13]  C. Scazzocchio,et al.  The genetic control of the molybdoflavoproteins in Aspergillus nidulans. IV. A comparison between purine hydroxylase I and II. , 1978, European journal of biochemistry.

[14]  C. Scazzocchio,et al.  A mutation defective in the xanthine alternative pathway of Aspergillus nidulans , 1978, Molecular and General Genetics MGG.

[15]  C. Scazzocchio,et al.  The uaY positive control gene of Aspergillus nidulans: fine structure, isolation of constitutive mutants and reversion patterns , 1991, Molecular and General Genetics MGG.

[16]  H. Arst,et al.  Cloning of the regulatory gene areA mediating nitrogen metabolite repression in Aspergillus nidulans. , 1986, The EMBO journal.

[17]  Meriel G. Jones,et al.  Characterization of nitrogen metabolite signalling in Aspergillus via the regulated degradation of areA mRNA , 2001, Molecular microbiology.

[18]  D. Cove,et al.  Nitrogen metabolite repression in Aspergillus nidulans , 1973, Molecular and General Genetics MGG.

[19]  C. Scazzocchio,et al.  Initiator constitutive mutation with an ‘up-promoter’ effect in Aspergillus nidulans , 1975, Nature.

[20]  A. Riggs,et al.  Genomic sequencing. , 1993, Methods in molecular biology.

[21]  J. Jurka,et al.  Rolling-circle transposons in eukaryotes , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[22]  M. Hood,et al.  Repetitive DNA in the automictic fungus Microbotryum violaceum , 2005, Genetica.

[23]  C. Scazzocchio The purine degradation pathway, genetics, biochemistry and regulation. , 1994, Progress in industrial microbiology.

[24]  C. Scazzocchio,et al.  Subtle hydrophobic interactions between the seventh residue of the zinc finger loop and the first base of an HGATAR sequence determine promoter‐specific recognition by the Aspergillus nidulans GATA factor AreA , 1997, The EMBO journal.

[25]  R. Hille,et al.  Xanthine oxidase and xanthine dehydrogenase , 1995, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[26]  C. Scazzocchio,et al.  A gene coding for the uric acid-xanthine permease of Aspergillus nidulans: inactivational cloning, characterization, and sequence of a cis-acting mutation. , 1989, Genetics.

[27]  C. Scazzocchio,et al.  Positive regulation in a eukaryote, a study of the uaY gene of Aspergillus nidulans: I. Characterization of alleles, dominance and complementation studies, and a fine structure map of the uaY--oxpA cluster. , 1982, Genetics.

[28]  R. Davies,et al.  The regulatory gene areA mediating nitrogen metabolite repression in Aspergillus nidulans. Mutations affecting specificity of gene activation alter a loop residue of a putative zinc finger. , 1990, The EMBO journal.

[29]  R. Hille The Mononuclear Molybdenum Enzymes. , 1997, Chemical reviews.

[30]  C. Scazzocchio,et al.  Double-joint PCR: a PCR-based molecular tool for gene manipulations in filamentous fungi. , 2004, Fungal genetics and biology : FG & B.

[31]  C. Scazzocchio The genetic control of molybdoflavoproteins in Aspergillus nidulans , 1973, Molecular and General Genetics MGG.

[32]  Robert P. Hausinger,et al.  Fe(II)/α-Ketoglutarate-Dependent Hydroxylases and Related Enzymes , 2004 .

[33]  A. Glatigny,et al.  Comparison of the sequences of the Aspergillus nidulans hxB and Drosophila melanogaster ma‐l genes with nifS from Azotobacter vinelandii suggests a mechanism for the insertion of the terminal sulphur atom in the molybdopterin cofactor , 2000, Molecular microbiology.

[34]  H. Arst A near terminal pericentric inversion leads to nitrogen metabolite derepression in aspergillus nidulans , 2004, Molecular and General Genetics MGG.

[35]  H. Arst,et al.  Genetic and Molecular Characterization of a Gene Encoding a Wide Specificity Purine Permease of Aspergillus nidulans Reveals a Novel Family of Transporters Conserved in Prokaryotes and Eukaryotes (*) , 1995, The Journal of Biological Chemistry.

[36]  Brandon S Gaut,et al.  Molecular and functional diversity of maize. , 2006, Current opinion in plant biology.

[37]  C. Scazzocchio,et al.  Biochemical and Genetical Studies of Purine Breakdown in Aspergillus , 1965, Nature.

[38]  R. Poulter,et al.  Vertebrate helentrons and other novel Helitrons. , 2003, Gene.

[39]  C. Scazzocchio,et al.  A single amino acid change in a pathway-specific transcription factor results in differing degrees of constitutivity, hyperinducibility and derepression of several structural genes. , 1995, Journal of molecular biology.

[40]  C. Weil,et al.  Cloning of the riboB locus of Aspergillus nidulans. , 1987, Gene.

[41]  Christina A. Cuomo,et al.  Sequencing of Aspergillus nidulans and comparative analysis with A. fumigatus and A. oryzae , 2005, Nature.

[42]  C. Scazzocchio,et al.  Sequence, regulation, and mutational analysis of the gene encoding urate oxidase in Aspergillus nidulans. , 1993, The Journal of biological chemistry.

[43]  C. Scazzocchio,et al.  The sequence and binding specificity of UaY, the specific regulator of the purine utilization pathway in Aspergillus nidulans, suggest an evolutionary relationship with the PPR1 protein of Saccharomyces cerevisiae. , 1995, The EMBO journal.

[44]  A. Glatigny,et al.  The hxB gene, necessary for the post‐translational activation of purine hydroxylases in Aspergillus nidulans, is independently controlled by the purine utilization and the nicotinate utilization transcriptional activating systems , 1999, Molecular microbiology.

[45]  C. Scazzocchio,et al.  The induction and repression of the enzymes of purine breakdown in Aspergillus nidulans. , 1968, Biochimica et biophysica acta.

[46]  D. Cove The induction and repression of nitrate reductase in the fungus Aspergillus nidulans. , 1966, Biochimica et biophysica acta.

[47]  C. Scazzocchio,et al.  The nature of an initiator constitutive mutation in Aspergillus nidulans , 1978, Nature.

[48]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[49]  C. Scazzocchio,et al.  Mutations in a dispensable region of the UaY transcription factor of Aspergillus nidulans differentially affect the expression of structural genes , 1997, Molecular microbiology.

[50]  H. Arst 13 – Formal Genetics and Molecular Biology of the Control of Gene Expression in Aspergillus nidulans , 1985 .

[51]  J. Doonan,et al.  Aspergillus nidulans contains a single actin gene which has unique intron locations and encodes a gamma-actin. , 1988, Gene.