Structural and functional analysis of Nro1/Ett1: a protein involved in translation termination in S. cerevisiae and in O2-mediated gene control in S. pombe

RNA. 2011 Jul;17(7):1213-24. doi: 10.1261/rna.2697111. Epub 2011 May 24.

Abstract

In Saccharomyces cerevisiae, the putative 2-OG-Fe(II) dioxygenase Tpa1 and its partner Ett1 have been shown to impact mRNA decay and translation. Hence, inactivation of these factors was shown to influence stop codon read-though. In addition, Tpa1 represses, by an unknown mechanism, genes regulated by Hap1, a transcription factor involved in the response to levels of heme and O(2). The Schizosaccharomyces pombe orthologs of Tpa1 and Ett1, Ofd1, and its partner Nro1, respectively, have been shown to regulate the stability of the Sre1 transcription factor in response to oxygen levels. To gain insight into the function of Nro1/Ett1, we have solved the crystal structure of the S. pombe Nro1 protein deleted of its 54 N-terminal residues. Nro1 unexpectedly adopts a Tetratrico Peptide Repeat (TPR) fold, a motif often responsible for protein or peptide binding. Two ligands, a sulfate ion and an unknown molecule, interact with a cluster of highly conserved amino acids on the protein surface. Mutation of these residues demonstrates that these ligand binding sites are essential for Ett1 function in S. cerevisiae, as investigated by assaying for efficient translation termination.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Codon, Terminator / genetics
  • Codon, Terminator / metabolism
  • Gene Expression Regulation, Fungal / drug effects*
  • Models, Molecular
  • Molecular Sequence Data
  • Multigene Family
  • Nuclear Proteins / chemistry*
  • Nuclear Proteins / physiology*
  • Oxygen / pharmacology*
  • Protein Biosynthesis*
  • Protein Folding
  • Protein Structure, Secondary
  • Protein Structure, Tertiary / genetics
  • Protein Structure, Tertiary / physiology
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / chemistry*
  • Saccharomyces cerevisiae Proteins / physiology*
  • Schizosaccharomyces / genetics*
  • Schizosaccharomyces / metabolism
  • Schizosaccharomyces pombe Proteins / chemistry*
  • Schizosaccharomyces pombe Proteins / physiology*
  • Sequence Homology, Amino Acid
  • Structure-Activity Relationship

Substances

  • Codon, Terminator
  • Ett1 protein, S cerevisiae
  • Nro1 protein, S pombe
  • Nuclear Proteins
  • Saccharomyces cerevisiae Proteins
  • Schizosaccharomyces pombe Proteins
  • Oxygen