Yoine Masato, Ohto Masa-aki, Onai Kiyoshi, Mita Satoru, Nakamura Kenzo
Laboratory of Biochemistry, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya 464-8601, Japan.
Plant J. 2006 Jul;47(1):49-62. doi: 10.1111/j.1365-313X.2006.02771.x. Epub 2006 Jun 1.
The low-beta-amylase1 (lba1) mutant of Arabidopsis thaliana has reduced sugar-induced expression of Atbeta-Amy and shows pleiotropic phenotypes such as early flowering; short day-sensitive growth; and seed germination that is hypersensitive to glucose and abscisic acid and resistant to mannose. lba1 was a missense mutation of UPF1 RNA helicase involved in nonsense-mediated mRNA decay (NMD), which eliminates mRNAs with premature termination codons (PTCs), and replaces highly conserved Gly851 of UPF1 with Glu. Expression of the wild-type UPF1 in lba1 rescued not only the reduced sugar-inducible gene expression, but also early flowering and altered seed-germination phenotypes. Sugar-inducible mRNAs were over-represented among transcripts decreased in sucrose-treated lba1 compared with Col plants, suggesting that UPF1 is involved in the expression of a subset of sugar-inducible genes. On the other hand, transcripts increased in lba1, which are likely to contain direct targets of NMD, included mRNAs for many transcription factors and metabolic enzymes that play diverse functions. Among these, the level of an alternatively spliced transcript of AtTFIIIA containing PTC was 17-fold higher in lba1 compared with Col plants, and it was reduced to the level in Col by expressing the wild-type UPF1. The lba1 mutant provides a good tool for studying NMD in plants.
拟南芥的低β-淀粉酶1(lba1)突变体具有降低的糖诱导Atbeta-Amy表达,并且表现出多效性表型,如早花;对短日照敏感的生长;以及对葡萄糖和脱落酸敏感且对甘露糖有抗性的种子萌发。lba1是参与无义介导的mRNA衰变(NMD)的UPF1 RNA解旋酶的错义突变,NMD可消除带有提前终止密码子(PTC)的mRNA,并将UPF1高度保守的Gly851替换为Glu。在lba1中表达野生型UPF1不仅挽救了糖诱导基因表达的降低,还挽救了早花和改变的种子萌发表型。与Col植物相比,在蔗糖处理的lba1中减少转录本中,糖诱导的mRNA占比过高,这表明UPF1参与了一部分糖诱导基因的表达。另一方面,在lba1中增加的转录本可能包含NMD的直接靶标,其中包括许多发挥不同功能的转录因子和代谢酶的mRNA。其中,含有PTC的AtTFIIIA可变剪接转录本在lba1中的水平比Col植物高17倍,通过表达野生型UPF1将其降低到Col中的水平。lba1突变体为研究植物中的NMD提供了一个很好的工具。