Translational regulation in response to changes in amino acid availability in Neurospora crassa

Mol Cell Biol. 1995 Oct;15(10):5235-45. doi: 10.1128/MCB.15.10.5235.

Abstract

We examined the regulation of Neurospora crassa arg-2 and cpc-1 in response to amino acid availability.arg-2 encodes the small subunit of arginine-specific carbamoyl phosphate synthetase; it is subject to unique negative regulation by Arg and is positively regulated in response to limitation for many different amino acids through a mechanism known as cross-pathway control. cpc-1 specifies a transcriptional activator important for crosspathway control. Expression of these genes was compared with that of the cytochrome oxidase subunit V gene, cox-5. Analyses of mRNA levels, polypeptide pulse-labeling results, and the distribution of mRNA in polysomes indicated that Arg-specific negative regulation of arg-2 affected the levels of both arg-2 mRNA and arg-2 mRNA translation. Negative translational effects on arg-2 and positive translational effects on cpc-1 were apparent soon after cells were provided with exogenous Arg. In cells limited for His, increased expression of arg-2 and cpc-1, and decreased expression of cox-5, also had translational and transcriptional components. The arg-2 and cpc-1 transcripts contain upstream open reading frames (uORFs), as do their Saccharomyces cerevisiae homologs CPA1 and GCN4. We examined the regulation of arg-2-lacZ reporter genes containing or lacking the uORF start codon; the capacity for arg-2 uORF translation appeared critical for controlling gene expression.

Publication types

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

MeSH terms

  • Amitrole / pharmacology
  • Arginine / pharmacology
  • Base Sequence
  • Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor*
  • Codon, Initiator
  • DNA-Binding Proteins / biosynthesis
  • DNA-Binding Proteins / genetics*
  • Electron Transport Complex IV / genetics
  • Fungal Proteins / biosynthesis
  • Fungal Proteins / genetics*
  • Gene Expression Regulation, Fungal / drug effects*
  • Genes, Fungal / genetics
  • Histidine / physiology
  • Hydro-Lyases / antagonists & inhibitors
  • Ligases / biosynthesis
  • Ligases / genetics*
  • Molecular Sequence Data
  • Neurospora crassa / enzymology
  • Neurospora crassa / genetics*
  • Open Reading Frames / genetics
  • Polyribosomes / metabolism
  • Protein Biosynthesis / drug effects*
  • RNA, Fungal / biosynthesis
  • RNA, Messenger / biosynthesis
  • Transcription, Genetic / drug effects

Substances

  • Codon, Initiator
  • DNA-Binding Proteins
  • Fungal Proteins
  • RNA, Fungal
  • RNA, Messenger
  • Histidine
  • Arginine
  • Electron Transport Complex IV
  • Hydro-Lyases
  • Ligases
  • Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor
  • carbamoyl phosphate synthetase (arginine-specific)
  • Amitrole