Laboratory of Plant Molecular Biology, Rockefeller University, New York, New York 10065, USA.
Plant Cell. 2009 Oct;21(10):3270-9. doi: 10.1105/tpc.109.070078. Epub 2009 Oct 23.
Eukaryotic processing bodies (P-bodies) are implicated in mRNA storage and mRNA decapping. We previously found that a decapping complex comprising Decapping 1 (DCP1), DCP2, and Varicose in Arabidopsis thaliana is essential for postembryonic development, but the underlying mechanism is poorly understood. Here, we characterized Arabidopsis DCP5, a homolog of human RNA-associated protein 55, as an additional P-body constituent. DCP5 associates with DCP1 and DCP2 and is required for mRNA decapping in vivo. In spite of its association with DCP2, DCP5 has no effect on DCP2 decapping activity in vitro, suggesting that the effect on decapping in vivo is indirect. In knockdown mutant dcp5-1, not only is mRNA decapping compromised, but the size of P-bodies is also significantly decreased. These results indicate that DCP5 is required for P-body formation, which likely facilitates efficient decapping. During wild-type seed germination, mRNAs encoding seed storage proteins (SSPs) are translationally repressed and degraded. By contrast, in dcp5-1, SSP mRNAs are translated, leading to accumulation of their products in germinated seedlings. In vitro experiments using wheat germ extracts confirmed that DCP5 is a translational repressor. Our results showed that DCP5 is required for translational repression and P-body formation and plays an indirect role in mRNA decapping.
真核生物加工体(P 体)参与 mRNA 储存和脱帽。我们之前发现拟南芥中包含脱帽酶 1(DCP1)、DCP2 和卷曲的脱帽复合物对于胚胎后发育是必不可少的,但其中的潜在机制尚不清楚。在这里,我们鉴定了拟南芥 DCP5 是人类 RNA 相关蛋白 55 的同源物,是另一种 P 体成分。DCP5 与 DCP1 和 DCP2 相关联,并且在体内需要进行 mRNA 脱帽。尽管与 DCP2 相关联,但 DCP5 对体外 DCP2 脱帽活性没有影响,这表明体内脱帽的影响是间接的。在 DCP5 敲低突变体 dcp5-1 中,不仅 mRNA 脱帽受到影响,而且 P 体的大小也显著减小。这些结果表明 DCP5 是 P 体形成所必需的,这可能有助于有效的脱帽。在野生型种子萌发过程中,编码种子贮藏蛋白(SSP)的 mRNAs 被翻译抑制并降解。相比之下,在 dcp5-1 中,SSP mRNAs 被翻译,导致其产物在萌发的幼苗中积累。使用小麦胚提取物进行的体外实验证实 DCP5 是一种翻译抑制剂。我们的结果表明 DCP5 对于翻译抑制和 P 体形成是必需的,并且在 mRNA 脱帽中发挥间接作用。