Yamashita Yui, Takamatsu Seidai, Glasbrenner Michael, Becker Thomas, Naito Satoshi, Beckmann Roland
Gene Center, Department of Biochemistry and Center for integrated Protein Science Munich (CiPSM), Ludwig-Maximilians-Universität München, Germany.
Graduate School of Life Science, Hokkaido University, Sapporo, Japan.
FEBS Lett. 2017 May;591(9):1266-1277. doi: 10.1002/1873-3468.12634. Epub 2017 Apr 10.
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.
拟南芥bZIP11是一种转录因子,在高蔗糖条件下调节氨基酸代谢。在翻译过程中,bZIP11的表达以蔗糖依赖的方式下调。先前的体内研究已确定第二个上游开放阅读框(uORF2)是bZIP11蔗糖依赖性翻译抑制的关键调控元件。然而,目前尚不清楚uORF2在高蔗糖条件下如何抑制bZIP11的表达。通过使用无细胞翻译系统的生化实验,我们报道了蔗糖介导的核糖体在uORF2终止密码子处的停滞。uORF2的C末端10个氨基酸(29-SFSVxFLxxLYYV-41)对核糖体停滞很重要。我们的结果表明,uORF2编码一种调节性新生肽,其功能是感知细胞内蔗糖丰度。这是细胞内蔗糖传感器的首次生化鉴定。