title: Sulfur-regulated control of the met-2⁺ gene of Neurospora crassa encoding cystathionine β-lyase.
abstract: BACKGROUND
  Cystathionine β-lyase performs an essential role in the transsulfuration pathway by its primary reaction of forming homocysteine from cystathionine. Understanding how the Neurospora crassa met-2⁺ gene, which encodes cystathionine β-lyase, is regulated is important in determining the basis of the cellular control of transsulfuration. The aim of this study was to determine the nature of a potential regulatory connection of met-2⁺ to the Neurospora sulfur regulatory network.
  
  
  RESULTS
  The cystathionine β-lyase (met-2⁺) gene was cloned by the identification of a cosmid genomic clone capable of transforming a met-2 mutant to methionine prototrophy and subsequently characterized. The gene contains a single intron and encodes a protein of 457 amino acids with conserved residues predicted to be important for catalysis and pyridoxal-5'-phosphate co-factor binding. The expression of met-2⁺ in wild-type N. crassa increased 3.1-fold under sulfur-limiting growth conditions as compared to the transcript levels seen under high sulfur growth conditions (i.e., repressing conditions). In a Δcys-3 strain, met-2⁺ transcript levels were substantially reduced under either low- or high-sulfur growth conditions. In addition, the presence of CYS3 activator binding sites on the met-2⁺ promoter was demonstrated by gel mobility shift assays.
  
  
  CONCLUSIONS
  In this report, we demonstrate the sulfur-regulated expression of the met-2⁺ gene and confirm its connection to the N. crassa sulfur regulatory circuit by the reduced expression observed in a Δcys-3 mutant and the in vitro detection of CYS3 binding sites in the met-2⁺ promoter. The data further adds to our understanding of the regulatory dynamics of transsulfuration.
authors.affiliation: Department of Biochemistry and Molecular Biology, Wright State University, Dayton, OH 45435, USA.
authors.forename: Brad S
authors.initials: BS
authors.lastname: Reveal
authors.forename: John V
authors.initials: JV
authors.lastname: Paietta
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D005800:Genes, Fungal; D008190:Lyases / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D009492:Neurospora crassa / Q000201:enzymology*; D013455:Sulfur / Q000378:metabolism*
chemical_list: D013455:Sulfur; D008190:Lyases; C019965:cystathionine beta-lyase