ASD4, a new GATA factor of Neurospora crassa, displays sequence-specific DNA binding and functions in ascus and ascospore development

Biochemistry. 2000 Sep 12;39(36):11065-73. doi: 10.1021/bi000886j.

Abstract

A new gene encoding a novel GATA factor, ASD4, of Neurospora crassa was isolated and demonstrated to possess one intron and to specify an open reading frame encoding a protein with 427 amino acid residues. The ASD4 protein contains a single GATA-type zinc finger and a putative coiled-coil domain. Unlike related proteins, DAL80 in yeast and NREB in Penicillium, ASD4 does not appear to be involved in regulation of nitrogen metabolism. An Asd-4 null mutant obtained by the rip procedure did not show any effect upon nitrogen control, but instead resulted in severe defects in ascus and ascospore genesis. The Asd-4 rip mutant is dominant to Asd-4+. A cross of the Asd-4 mutant with wild-type resulted in fruiting bodies that appeared to be normal macroscopically but which were complete devoid of asci and ascospores. Introduction of the Asd-4+ gene into the Asd-4 rip mutant corrected the defect in ascus and ascospore development in crosses with wild-type. Mobility shift assays demonstrated that ASD4 acts as a sequence-specific DNA binding protein and recognizes DNA fragments that contain GATA core elements. Gel filtration and cross-linking experiments revealed that the ASD4 protein exists as a tetramer in solution. These results suggest that the ASD4 protein functions positively as a transcriptional regulator of sexual development in Neurospora.

Publication types

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

MeSH terms

  • Blotting, Northern
  • Cysteine / chemistry
  • Cysteine / genetics
  • Cysteine / physiology
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / isolation & purification*
  • DNA-Binding Proteins / physiology*
  • Fungal Proteins / chemistry
  • Fungal Proteins / genetics
  • Fungal Proteins / isolation & purification*
  • Fungal Proteins / physiology*
  • Mutagenesis
  • Neurospora crassa / chemistry*
  • Neurospora crassa / genetics
  • Neurospora crassa / physiology*
  • Nitrogen / metabolism
  • Protein Structure, Tertiary / genetics
  • Repetitive Sequences, Amino Acid
  • Solutions
  • Spores, Fungal / chemistry
  • Spores, Fungal / genetics
  • Spores, Fungal / physiology
  • Transformation, Genetic
  • Zinc Fingers* / genetics
  • Zinc Fingers* / physiology

Substances

  • DNA-Binding Proteins
  • Fungal Proteins
  • Solutions
  • Cysteine
  • Nitrogen