FfVel1 and FfLae1, components of a velvet‐like complex in Fusarium fujikuroi, affect differentiation, secondary metabolism and virulence

Besides industrially produced gibberellins (GAs), Fusarium fujikuroi is able to produce additional secondary metabolites such as the pigments bikaverin and neurosporaxanthin and the mycotoxins fumonisins and fusarin C. The global regulation of these biosynthetic pathways is only poorly understood. Recently, the velvet complex containing VeA and several other regulatory proteins was shown to be involved in global regulation of secondary metabolism and differentiation in Aspergillus nidulans. Here, we report on the characterization of two components of the F. fujikuroi velvet‐like complex, FfVel1 and FfLae1. The gene encoding this first reported LaeA orthologue outside the class of Eurotiomycetidae is upregulated in ΔFfvel1 microarray‐studies and FfLae1 interacts with FfVel1 in the nucleus. Deletion of Ffvel1 and Fflae1 revealed for the first time that velvet can simultaneously act as positive (GAs, fumonisins and fusarin C) and negative (bikaverin) regulator of secondary metabolism, and that both components affect conidiation and virulence of F. fujikuroi. Furthermore, the velvet‐like protein FfVel2 revealed similar functions regarding conidiation, secondary metabolism and virulence as FfVel1. Cross‐genus complementation studies of velvet complex component mutants between Fusarium, Aspergillus and Penicillium support an ancient origin for this complex, which has undergone a divergence in specific functions mediating development and secondary metabolism.

[1]  Birgit Hoff,et al.  Two Components of a velvet-Like Complex Control Hyphal Morphogenesis, Conidiophore Development, and Penicillin Biosynthesis in Penicillium chrysogenum , 2010, Eukaryotic Cell.

[2]  C. Scazzocchio,et al.  Heterochromatic marks are associated with the repression of secondary metabolism clusters in Aspergillus nidulans , 2010, Molecular microbiology.

[3]  M. Busman,et al.  FvVE1 regulates biosynthesis of the mycotoxins fumonisins and fusarins in Fusarium verticillioides. , 2009, Journal of agricultural and food chemistry.

[4]  H. Humpf,et al.  Biosynthesis of the red pigment bikaverin in Fusarium fujikuroi: genes, their function and regulation , 2009, Molecular microbiology.

[5]  N. Keller,et al.  Distinct Roles for VeA and LaeA in Development and Pathogenesis of Aspergillus flavus , 2009, Eukaryotic Cell.

[6]  W. K. Wilson,et al.  Protostadienol biosynthesis and metabolism in the pathogenic fungus Aspergillus fumigatus. , 2009, Organic letters.

[7]  Carlos García-Estrada,et al.  The global regulator LaeA controls penicillin biosynthesis, pigmentation and sporulation, but not roquefortine C synthesis in Penicillium chrysogenum. , 2009, Biochimie.

[8]  P. Hedden,et al.  Loss of Gibberellin Production in Fusarium verticillioides (Gibberella fujikuroi MP-A) Is Due to a Deletion in the Gibberellic Acid Gene Cluster , 2008, Applied and Environmental Microbiology.

[9]  J. Frisvad,et al.  Requirement of LaeA for secondary metabolism and sclerotial production in Aspergillus flavus. , 2008, Fungal genetics and biology : FG & B.

[10]  Harald Berger,et al.  Dissecting individual steps of nitrogen transcription factor cooperation in the Aspergillus nidulans nitrate cluster , 2008, Molecular microbiology.

[11]  B. Tudzynski,et al.  Cross-Species Hybridization with Fusarium verticillioides Microarrays Reveals New Insights into Fusarium fujikuroi Nitrogen Regulation and the Role of AreA and NMR , 2008, Eukaryotic Cell.

[12]  A. M. Calvo.  The VeA regulatory system and its role in morphological and chemical development in fungi. , 2008, Fungal genetics and biology : FG & B.

[13]  G. Braus,et al.  Animal Versus Wind Dispersal and the Robustness of Tree Species to Deforestation , 2008, Science.

[14]  C. Woloshuk,et al.  Role of AREA, a regulator of nitrogen metabolism, during colonization of maize kernels and fumonisin biosynthesis in Fusarium verticillioides. , 2008, Fungal genetics and biology : FG & B.

[15]  Kap-Hoon Han,et al.  The Aspergillus nidulans esdC (early sexual development) gene is necessary for sexual development and is controlled by veA and a heterotrimeric G protein. , 2008, Fungal genetics and biology : FG & B.

[16]  G. Braus,et al.  Neurospora crassa ve-1 affects asexual conidiation. , 2008, Fungal genetics and biology : FG & B.

[17]  R. Fischer,et al.  Mapping the interaction sites of Aspergillus nidulans phytochrome FphA with the global regulator VeA and the White Collar protein LreB , 2008, Molecular Genetics and Genomics.

[18]  D. Hoffmeister,et al.  Accurate prediction of the Aspergillus nidulans terrequinone gene cluster boundaries using the transcriptional regulator LaeA. , 2007, Fungal genetics and biology : FG & B.

[19]  Paul Horton,et al.  Nucleic Acids Research Advance Access published May 21, 2007 WoLF PSORT: protein localization predictor , 2007 .

[20]  M. Busman,et al.  The Fusarium verticillioides FUM Gene Cluster Encodes a Zn(II)2Cys6 Protein That Affects FUM Gene Expression and Fumonisin Production , 2007, Eukaryotic Cell.

[21]  Jennifer R Wortman,et al.  Transcriptional Regulation of Chemical Diversity in Aspergillus fumigatus by LaeA , 2007, PLoS pathogens.

[22]  U. Kück,et al.  A Homologue of the Aspergillus velvet Gene Regulates both Cephalosporin C Biosynthesis and Hyphal Fragmentation in Acremonium chrysogenum , 2007, Applied and Environmental Microbiology.

[23]  D. Hoffmeister,et al.  Natural products of filamentous fungi: enzymes, genes, and their regulation. , 2007, Natural product reports.

[24]  A. Brakhage,et al.  The light-dependent regulator velvet A of Aspergillus nidulans acts as a repressor of the penicillin biosynthesis , 2007, Archives of Microbiology.

[25]  E. Espeso,et al.  Aspergillus nidulans VeA subcellular localization is dependent on the importin α carrier and on light , 2007, Molecular microbiology.

[26]  A. M. Calvo,et al.  Production of cyclopiazonic acid, aflatrem, and aflatoxin by Aspergillus flavus is regulated by veA, a gene necessary for sclerotial formation , 2007, Applied Microbiology and Biotechnology.

[27]  Robert H Schiestl,et al.  Quick and easy yeast transformation using the LiAc/SS carrier DNA/PEG method , 2007, Nature Protocols.

[28]  H. Arst.  Faculty Opinions recommendation of Aspergillus nidulans VeA subcellular localization is dependent on the importin alpha carrier and on light. , 2006 .

[29]  Shaojie Li,et al.  FvVE1 regulates filamentous growth, the ratio of microconidia to macroconidia and cell wall formation in Fusarium verticillioides , 2006, Molecular microbiology.

[30]  D. Noordermeer,et al.  Secondary metabolic gene cluster silencing in Aspergillus nidulans , 2006, Molecular microbiology.

[31]  Christopher M. Crew,et al.  A high-throughput gene knockout procedure for Neurospora reveals functions for multiple transcription factors , 2006, Proceedings of the National Academy of Sciences.

[32]  R. Baltensperger,et al.  FiRe and microarrays: a fast answer to burning questions. , 2006, Trends in plant science.

[33]  Jan Löwe,et al.  RF cloning: a restriction-free method for inserting target genes into plasmids. , 2006, Journal of biochemical and biophysical methods.

[34]  K. Turksen,et al.  Isolation and characterization , 2006 .

[35]  Nancy P Keller,et al.  Genomic mining for Aspergillus natural products. , 2006, Chemistry & biology.

[36]  T. Utterback,et al.  Comparative analysis of 87,000 expressed sequence tags from the fumonisin-producing fungus Fusarium verticillioides. , 2005, Fungal genetics and biology : FG & B.

[37]  J. Frisvad,et al.  LaeA, a Regulator of Morphogenetic Fungal Virulence Factors , 2005, Eukaryotic Cell.

[38]  G. Braus,et al.  Deletion and Allelic Exchange of the Aspergillus fumigatus veA Locus via a Novel Recyclable Marker Module , 2005, Eukaryotic Cell.

[39]  M. Ferguson,et al.  A simple and rapid technique for fluorescence staining of fungal nuclei , 1984, Current Microbiology.

[40]  B. Tudzynski.  Gibberellin biosynthesis in fungi: genes, enzymes, evolution, and impact on biotechnology , 2005, Applied Microbiology and Biotechnology.

[41]  U. Kück,et al.  Use of bimolecular fluorescence complementation to demonstrate transcription factor interaction in nuclei of living cells from the filamentous fungus Acremonium chrysogenum , 2005, Current Genetics.

[42]  N. Keller,et al.  Endogenous Lipogenic Regulators of Spore Balance in Aspergillus nidulans , 2004, Eukaryotic Cell.

[43]  S. Richter,et al.  Deletion of the Gibberella fujikuroi glutamine synthetase gene has significant impact on transcriptional control of primary and secondary metabolism , 2004, Molecular microbiology.

[44]  A. M. Calvo,et al.  veA Is Required for Toxin and Sclerotial Production in Aspergillus parasiticus , 2004, Applied and Environmental Microbiology.

[45]  Jeong-Ah Seo,et al.  Discontinuous distribution of fumonisin biosynthetic genes in the Gibberella fujikuroi species complex. , 2004, Mycological research.

[46]  P. Hedden,et al.  The NADPH-cytochrome P450 Reductase Gene from Gibberella fujikuroi Is Essential for Gibberellin Biosynthesis* , 2004, Journal of Biological Chemistry.

[47]  J. Bok,et al.  LaeA, a Regulator of Secondary Metabolism in Aspergillus spp , 2004, Eukaryotic Cell.

[48]  A. M. Calvo,et al.  The Expression of Sterigmatocystin and Penicillin Genes in Aspergillus nidulans Is Controlled by veA, a Gene Required for Sexual Development , 2003, Eukaryotic Cell.

[49]  J. Flaherty,et al.  PAC1, a pH-Regulatory Gene from Fusarium verticillioides , 2003, Applied and Environmental Microbiology.

[50]  P. Hedden,et al.  Characterization of the Final Two Genes of the Gibberellin Biosynthesis Gene Cluster of Gibberella fujikuroi , 2003, Journal of Biological Chemistry.

[51]  S. Chae,et al.  Identification and Cloning of jipA Encoding a Polypeptide That Interacts with a Homolog of Yeast Rad6, UVSJ in Aspergillus nidulans , 2003 .

[52]  Daren W Brown,et al.  Co-expression of 15 contiguous genes delineates a fumonisin biosynthetic gene cluster in Gibberella moniliformis. , 2003, Fungal genetics and biology : FG & B.

[53]  B. Tudzynski,et al.  AREA directly mediates nitrogen regulation of gibberellin biosynthesis in Gibberella fujikuroi, but its activity is not affected by NMR , 2003, Molecular microbiology.

[54]  U. Kück,et al.  Versatile EGFP reporter plasmids for cellular localization of recombinant gene products in filamentous fungi , 2003, Current Genetics.

[55]  Kyung Kim,et al.  The veA gene activates sexual development in Aspergillus nidulans. , 2002, Fungal genetics and biology : FG & B.

[56]  U. Kück,et al.  Functional analysis of the C6 zinc finger gene pro1 involved in fungal sexual development. , 2002, Fungal genetics and biology : FG & B.

[57]  E. Coppin.  The fle1 gene encoding a C2H2 zinc finger protein co‐ordinates male and female sexual differentiation in Podospora anserina , 2002, Molecular microbiology.

[58]  C. Maragos,et al.  Fusarium Species from Nepalese Rice and Production of Mycotoxins and Gibberellic Acid by Selected Species , 2000, Applied and Environmental Microbiology.

[59]  Ogas.  Gibberellins , 2000, Current Biology.

[60]  G. Marzluf,et al.  Isolation, characterization and disruption of the areA nitrogen regulatory gene of Gibberella fujikuroi , 1999, Molecular and General Genetics MGG.

[61]  B. Tudzynski,et al.  Gibberellin biosynthetic pathway in Gibberella fujikuroi: evidence for a gene cluster. , 1998, Fungal genetics and biology : FG & B.

[62]  H. Kawaide,et al.  Gibberellin biosynthesis in Gibberella fujikuroi: cloning and characterization of the copalyl diphosphate synthase gene , 1998, Current Genetics.

[63]  B. Regenberg,et al.  Dip5p mediates high-affinity and high-capacity transport of L-glutamate and L-aspartate in Saccharomyces cerevisiae , 1998, Current Genetics.

[64]  T. Adams,et al.  Asexual Sporulation in Aspergillus nidulans , 1998, Microbiology and Molecular Biology Reviews.

[65]  S. McCormick,et al.  The TRI11 Gene of Fusarium sporotrichioides Encodes a Cytochrome P-450 Monooxygenase Required for C-15 Hydroxylation in Trichothecene Biosynthesis , 1998, Applied and Environmental Microbiology.

[66]  D. Weisleder,et al.  Analytical determination of fumonisins and other metabolites produced by Fusarium moniliforme and related species on corn. , 1996, Advances in experimental medicine and biology.

[67]  Fred Winston,et al.  Construction of a set of convenient saccharomyces cerevisiae strains that are isogenic to S288C , 1995, Yeast.

[68]  J. Becker,et al.  Isolation and characterization of a Saccharomyces cerevisiae peptide transport gene , 1994, Molecular and cellular biology.

[69]  A. Riggs,et al.  Genomic Sequencing , 2010 .

[70]  J. Cenis,et al.  Rapid extraction of fungal DNA for PCR amplification. , 1992, Nucleic acids research.

[71]  M. Dante,et al.  Multifunctional yeast high-copy-number shuttle vectors. , 1992, Gene.

[72]  J. E. Oltra,et al.  Fusarin C and 8Z-fusarin C from Gibberella fujikuroi , 1991 .

[73]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[74]  J. Doyle,et al.  Isolation of plant DNA from fresh tissue , 1990 .

[75]  E. Selker,et al.  Use of a bacterial Hygromycin B resistance gene as a dominant selectable marker in Neurospora crassa , 1989 .

[76]  J. Leslie,et al.  Nitrate reduction mutants of fusarium moniliforme (gibberella fujikuroi). , 1988, Genetics.

[77]  S. Rogers,et al.  Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis. , 1986, Science.

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

[79]  J. Davies,et al.  Plasmid-encoded hygromycin B resistance: the sequence of hygromycin B phosphotransferase gene and its expression in Escherichia coli and Saccharomyces cerevisiae. , 1983, Gene.

[80]  J. Bu’lock,et al.  Bikaverin and norbikaverin, benzoxanthentrione pigments of Gibberella fujikuroi. , 1971, Journal of the Chemical Society. Perkin transactions 1.

[81]  G. Cragg,et al.  Studies on the biosynthesis of gibberellins from (−)-kaurenoic acid in cultures of Gibberella Fujikuroi , 1966 .

[82]  M A DARKEN,et al.  Production of gibberellic acid by fermentation. , 1959, Applied microbiology.

[83]  K. D. Macdonald,et al.  The genetics of Aspergillus nidulans. , 1953, Advances in genetics.