Molecular characterization of mutants of the acetate regulatory gene facB of Aspergillus nidulans

Fungal Genet Biol. 1997 Oct;22(2):92-102. doi: 10.1006/fgbi.1997.1007.

Abstract

The facB gene of Aspergillus nidulans encodes a DNA binding transcriptional activator required for growth on acetate as a sole carbon source. FacB contains N-terminal GAL4-like Zn(II)2Cys6 (or C6 zinc) binuclear cluster DNA binding and leucine zipper-like heptad repeat motifs and central and C-terminal acidic alpha-helical regions. facB recessive loss of function mutants are deficient in acetate induction of acetyl-CoA synthase, isocitrate lyase, malate synthase, acetamidase, and NADP-isocitrate dehydrogenase. Characterization of lesions in facB mutant alleles has localized important functional regions of the FacB protein. Two extreme mutants are shown to lack the C-terminal region of the protein. Two temperature sensitive mutants contain amino acid substitutions in the DNA binding domain and are shown to affect acetate induction of amdS-lacZ expression and confer temperature sensitive in vitro DNA binding. Two temperature sensitive facB mutations result in thermolability of acetyl-CoA synthase, isocitrate lyase, and malate synthase but not acetamidase or NADP-isocitrate dehydrogenase in crude extracts. This suggests that FacB may have a structural role in acetate metabolism in addition to its regulatory function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetates / metabolism*
  • Alleles
  • Amidohydrolases / genetics
  • Amidohydrolases / metabolism
  • Amino Acid Sequence
  • Aspergillus nidulans / enzymology
  • Aspergillus nidulans / genetics*
  • Aspergillus nidulans / metabolism
  • Enzyme Induction
  • Enzyme Stability
  • Escherichia coli / genetics
  • Fungal Proteins / biosynthesis
  • Fungal Proteins / metabolism
  • Genes, Fungal / genetics
  • Molecular Sequence Data
  • Mutation / genetics*
  • Mutation / physiology
  • Recombinant Fusion Proteins
  • Sequence Analysis, DNA
  • Sequence Deletion / genetics
  • Temperature
  • Trans-Activators / genetics*
  • Trans-Activators / metabolism
  • Zinc Fingers

Substances

  • Acetates
  • Fungal Proteins
  • Recombinant Fusion Proteins
  • Trans-Activators
  • Amidohydrolases
  • acetamidase