Molecular Cloning and Analysis of Nre, the Major Nitrogen Regulatory Gene of Penicillium Chrysogenum

Curr Genet. 1995 Jan;27(2):150-8. doi: 10.1007/BF00313429.

Abstract

We have isolated the Penicillium chrysogenum nre gene which is homologous to the major nitrogen regulatory genes areA from Aspergillus nidulans and nit-2 from Neurospora crassa. Overall, nre shows 60% identity to areA and 30% identity to nit-2 at the amino-acid level. The gene encodes a protein of 835 amino-acid residues and contains a single Cys2/Cys2-type zinc finger with an adjacent basic region and a putative acidic activation region. In the DNA-binding domain, 98% of the amino-acid residues are identical in nre, areA and nit-2. The nre gene has been shown to be functional in N. crassa by heterologous complementation of a nit-2 mutant. Growth tests indicated that transformants could utilize nitrate, amino-acids, purines and amides as sole nitrogen sources. Nitrate reductase activity assays performed with transformants demonstrated that nitrogen control was completely normal. Complementation of N. crassa nit-2 mutants with 5'-deletion clones of nre suggests the possible presence of an internal promoter within the coding region. Northern analysis and ribonuclease protection assays of total cellular RNA indicated that nre encodes a 3.2-kb transcript which is reduced in content under conditions of nitrogen repression.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Aspergillus nidulans / genetics
  • Base Sequence
  • Cloning, Molecular
  • Fungal Proteins / biosynthesis
  • Fungal Proteins / genetics*
  • Fungal Proteins / metabolism
  • Gene Expression Regulation, Fungal*
  • Genes, Fungal*
  • Molecular Sequence Data
  • Mutation
  • Neurospora crassa / genetics
  • Neurospora crassa / metabolism
  • Nitrate Reductases / metabolism
  • Nitrogen / metabolism*
  • Penicillium chrysogenum / genetics*
  • Restriction Mapping
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • Transcription Factors / biosynthesis
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Zinc Fingers

Substances

  • AREA protein, Penicillium chrysogenum
  • Fungal Proteins
  • Transcription Factors
  • Nitrate Reductases
  • Nitrogen

Associated data

  • GENBANK/U02612