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Identification of Arabidopsis thaliana upstream open reading frames encoding peptide sequences that cause ribosomal arrest

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Title: Identification of Arabidopsis thaliana upstream open reading frames encoding peptide sequences that cause ribosomal arrest
Authors: Hayashi, Noriya Browse this author
Sasaki, Shun Browse this author
Takahashi, Hiro Browse this author
Yamashita, Yui Browse this author
Naito, Satoshi Browse this author →KAKEN DB
Onouchi, Hitoshi Browse this author
Keywords: arabidopsis
codon nucleotides
open reading frames
Issue Date: 6-Sep-2017
Publisher: Oxford University Press
Journal Title: Nucleic acids research
Volume: 45
Issue: 15
Start Page: 8844
End Page: 8858
Publisher DOI: 10.1093/nar/gkx528
PMID: 28637336
Abstract: Specific sequences of certain nascent peptides cause programmed ribosomal arrest during mRNA translation to control gene expression. In eukaryotes, most known regulatory arrest peptides are encoded by upstream open reading frames (uORFs) present in the 5'-untranslated region ofmRNAs. However, to date, a limited number of eukaryotic uORFs encoding arrest peptides have been reported. Here, we searched for arrest peptide-encoding uORFs among Arabidopsis thaliana uORFs with evolutionarily conserved peptide sequences. Analysis of in vitro translation products of 22 conserved uORFs identified three novel uORFs causing ribosomal arrest in a peptide sequence-dependent manner. Stop codon-scanning mutagenesis, in which the effect of changing the uORF stop codon position on the ribosomal arrest was examined, and toeprint analysis revealed that two of the three uORFs cause ribosomal arrest during translation elongation, whereas the other one causes ribosomal arrest during translation termination. Transient expression assays showed that the newly identified arrest-causing uORFs exerted a strong sequence-dependent repressive effect on the expression of the downstream reporter gene in A. thaliana protoplasts. These results suggest that the peptide sequences of the three uORFs identified in this study cause ribosomal arrest in the uORFs, thereby repressing the expression of proteins encoded by the main ORFs.
Type: article
Appears in Collections:農学院・農学研究院 (Graduate School of Agriculture / Faculty of Agriculture) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

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