Molecular cloning and analysis of nre, the major nitrogen regulatory gene of Penicillium chrysogenum

• Published in: Current genetics 1995, Vol.27 (2), p.150-158
• Main Authors: Haas, H, Bauer, B, Redl, B, Stoeffler, G, Marzluf, G.A. (Ohio State Univ., Columbus (USA). Dept. of Biochemistry)
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Aspergillus nidulans - genetics; Base Sequence; CLONACION MOLECULAR; CLONAGE MOLECULAIRE; Cloning; Cloning, Molecular; Fungal Proteins - biosynthesis; Fungal Proteins - genetics; Fungal Proteins - metabolism; GENE; Gene Expression Regulation, Fungal; GENES; Genes, Fungal; METABOLISME DE L'AZOTE; METABOLISMO DEL NITROGENO; MOLECULAR CLONING; Molecular Sequence Data; Mutation; Neurospora crassa - genetics; Neurospora crassa - metabolism; Nitrate Reductases - metabolism; NITROGEN METABOLISM; Nitrogen regulation; PENICILLIUM; Penicillium chrysogenum; Penicillium chrysogenum - genetics; Pflanzenzuechtung; POLIMERIZACION; Polymerase chain reaction; POLYMERISATION; POLYMERIZATION; PROTEINAS; PROTEINE; PROTEINS; Restriction Mapping; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Transcription Factors - biosynthesis; Transcription Factors - genetics; Transcription Factors - metabolism; Zinc Fingers
• ISSN: 0172-8083; 1432-0983
• DOI: 10.1007/BF00313429

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.