Ribosome stalling is responsible for arginine-specific translational attenuation in Neurospora crassa.
The Neurospora crassa arg-2 upstream open reading frame (uORF) plays a role in negative arginine-specific translational regulation. Primer extension inhibition analyses of arg-2 uORF-containing RNA translated in a cell-free system in which arginine-specific regulation was retained revealed "toeprints" corresponding to ribosomes positioned at the uORF initiation and termination codons and at the downstream initiation codon. At high arginine concentrations, the toeprint signal corresponding to ribosomes at the uORF termination codon rapidly increased; a new, broad toeprint that represents additional ribosomes stalled on the uORF appeared 21 to 30 nucleotides upstream of this site; and the toeprint signal corresponding to ribosomes at the downstream initiation codon decreased. These data suggest that arginine increases ribosomal stalling and thereby decreases translation from the downstream initiation codon.
Wang
Z
Z
Department of Biochemistry and Molecular Biology, Oregon Graduate Institute of Science & Technology, Portland 97291-1000, USA.
Sachs
M S
MS
D000595:Amino Acid Sequence; D001120:Arginine / Q000096:biosynthesis / Q000502:physiology*; D001483:Base Sequence; D019733:Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor; D003513:Cycloheximide / Q000494:pharmacology; D017930:Genes, Reporter; D008025:Ligases / Q000235:genetics*; D008156:Luciferases / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D009492:Neurospora crassa / Q000201:enzymology / Q000235:genetics*; D016366:Open Reading Frames; D014176:Protein Biosynthesis; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011691:Puromycin / Q000494:pharmacology; D012313:RNA / Q000378:metabolism; D012270:Ribosomes / Q000201:enzymology* / Q000235:genetics
D011500:Protein Synthesis Inhibitors; D011691:Puromycin; D012313:RNA; D001120:Arginine; D003513:Cycloheximide; D008156:Luciferases; D008025:Ligases; D019733:Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor; C064716:carbamoyl phosphate synthetase (arginine-specific)