Isolation, characterization and disruption of the areA nitrogen regulatory gene of Gibberella fujikuroi

Mol Gen Genet. 1999 Feb;261(1):106-14. doi: 10.1007/s004380050947.

Abstract

The gene areA-GF, a homologue of the major nitrogen regulatory genes nit-2, areA, nre and NUT1 of Neurospora crassa, Aspergillus nidulans, Penicillium chrysogenum and Magnaporthe grisea, respectively, was cloned from the gibberellin (GA)-producing rice pathogen Gibberella fujikuroi. areA-GF encodes a protein of 972 amino acid residues which contains a single putative zinc finger DNA-binding domain that is at least 98% identical to the zinc finger domains of the homologous fungal proteins. The areA-GF gene has been shown to be functional in N. crassa by heterologous complementation of a RIP induced nit-2 mutant. The transformation rate was nearly as high as in a homologous complementation control. Transformants were able to utilize nitrate and expressed a normally regulated nitrate reductase activity. To generate areA-GF- mutants, gene replacement experiments were performed using a linearized replacement vector carrying the hygromycin B phosphotransferase (hph) gene. The replacement of the zinc finger by the hygromycin cassette resulted in transformants which were unable to utilize nitrogen sources other than ammonium and glutamine, and gave significantly reduced gibberellin production yields. Complementation of such a mutant with the wild-type gene led to the full recovery of gibberellin production.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Cloning, Molecular
  • DNA, Fungal / chemistry
  • DNA, Fungal / genetics
  • DNA, Fungal / isolation & purification*
  • DNA-Binding Proteins / genetics
  • Fungal Proteins / genetics*
  • Fungal Proteins / physiology
  • Gene Expression Regulation, Fungal
  • Gene Targeting
  • Genes, Fungal / genetics*
  • Genetic Complementation Test
  • Gibberella / genetics*
  • Gibberellins / biosynthesis
  • Molecular Sequence Data
  • Mutation
  • Neurospora crassa / genetics
  • Nitrate Reductase
  • Nitrate Reductases / genetics
  • Nitrogen / metabolism
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • Transcription Factors / genetics*
  • Transcription Factors / physiology

Substances

  • AreA protein, Aspergillus nidulans
  • DNA, Fungal
  • DNA-Binding Proteins
  • Fungal Proteins
  • Gibberellins
  • NIT2 protein, Neurospora crassa
  • Transcription Factors
  • Nitrate Reductases
  • Nitrate Reductase
  • Nitrogen

Associated data

  • GENBANK/Y11006