title: Sucrose sensing through nascent peptide-meditated ribosome stalling at the stop codon of Arabidopsis bZIP11 uORF2.
abstract: Arabidopsis bZIP11 is a transcription factor that modulates amino acid metabolism under high-sucrose conditions. Expression of bZIP11 is downregulated in a sucrose-dependent manner during translation. Previous in vivo studies have identified the second upstream open reading frame (uORF2) as an essential regulatory element for the sucrose-dependent translational repression of bZIP11. However, it remains unclear how uORF2 represses bZIP11 expression under high-sucrose conditions. Through biochemical experiments using cell-free translation systems, we report on sucrose-mediated ribosome stalling at the stop codon of uORF2. The C-terminal 10 amino acids (29-SFSVxFLxxLYYV-41) of uORF2 are important for ribosome stalling. Our results demonstrate that uORF2 encodes a regulatory nascent peptide that functions to sense intracellular sucrose abundance. This is the first biochemical identification of the intracellular sucrose sensor.
authors.affiliation: Gene Center, Department of Biochemistry and Center for integrated Protein Science Munich (CiPSM), Ludwig-Maximilians-Universität München, Germany.
authors.forename: Yui
authors.initials: Y
authors.lastname: Yamashita
authors.affiliation: Graduate School of Life Science, Hokkaido University, Sapporo, Japan.
authors.forename: Seidai
authors.initials: S
authors.lastname: Takamatsu
authors.affiliation: Gene Center, Department of Biochemistry and Center for integrated Protein Science Munich (CiPSM), Ludwig-Maximilians-Universität München, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Glasbrenner
authors.affiliation: Gene Center, Department of Biochemistry and Center for integrated Protein Science Munich (CiPSM), Ludwig-Maximilians-Universität München, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Becker
authors.affiliation: Graduate School of Life Science, Hokkaido University, Sapporo, Japan.|Graduate School of Agriculture, Hokkaido University, Sapporo, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Naito
authors.affiliation: Gene Center, Department of Biochemistry and Center for integrated Protein Science Munich (CiPSM), Ludwig-Maximilians-Universität München, Germany.
authors.forename: Roland
authors.initials: R
authors.lastname: Beckmann
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D050976:Basic-Leucine Zipper Transcription Factors / Q000235:genetics* / Q000378:metabolism; D002474:Cell-Free System / Q000378:metabolism; D018388:Codon, Terminator / Q000235:genetics*; D018506:Gene Expression Regulation, Plant; D015151:Immunoblotting; D009154:Mutation; D016366:Open Reading Frames / Q000235:genetics*; D010455:Peptides / Q000235:genetics* / Q000378:metabolism; D014176:Protein Biosynthesis / Q000235:genetics; D012270:Ribosomes / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013395:Sucrose / Q000378:metabolism*
chemical_list: C112599:ATB2 protein, Arabidopsis; D029681:Arabidopsis Proteins; D050976:Basic-Leucine Zipper Transcription Factors; D018388:Codon, Terminator; D010455:Peptides; D013395:Sucrose
keywords: in vitro translation; nascent peptide; ribosome stalling; sucrose sensing; upstream ORF