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FEBS Lett. 2017 May;591(9):1266-1277. doi: 10.1002/1873-3468.12634. Epub 2017 Apr 10.

Sucrose sensing through nascent peptide-meditated ribosome stalling at the stop codon of Arabidopsis bZIP11 uORF2.

Author information

1
Gene Center, Department of Biochemistry and Center for integrated Protein Science Munich (CiPSM), Ludwig-Maximilians-Universität München, Germany.
2
Graduate School of Life Science, Hokkaido University, Sapporo, Japan.
3
Graduate School of Agriculture, Hokkaido University, Sapporo, Japan.

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.

KEYWORDS:

in vitro translation; nascent peptide; ribosome stalling; sucrose sensing; upstream ORF

PMID:
28369795
DOI:
10.1002/1873-3468.12634
[Indexed for MEDLINE]
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