Ramirez Carmen Velasco, Vilela Cristina, Berthelot Karine, McCarthy John E G
Posttranscriptional Control Group, Department of Biomolecular Sciences, University of Manchester Institute of Science and Technology (UMIST), P.O. Box 88, M60 1QD, UK.
J Mol Biol. 2002 May 10;318(4):951-62. doi: 10.1016/S0022-2836(02)00162-6.
Decapping by Dcp1 in Saccharomyces cerevisiae is a key step in mRNA degradation. However, the cap also binds the eukaryotic initiation factor (eIF) complex 4F and its associated proteins. Characterisation of the relationship between decapping and interactions involving eIF4F is an essential step towards understanding polysome disassembly and mRNA decay. Three types of observation suggest how changes in the functional status of eIF4F modulate mRNA stability in vivo. First, partial disruption of the interaction between eIF4E and eIF4G, caused by mutations in eIF4E or the presence of the yeast 4E-binding protein p20, stabilised mRNAs. The interactions of eIF4G and p20 with eIF4E may therefore act to modulate the decapping process. Since we also show that the in vitro decapping rate is not directly affected by the nature of the body of the mRNA, this suggests that changes in eIF4F structure could play a role in triggering decapping during mRNA decay. Second, these effects were seen in the absence of extreme changes in global translation rates in the cell, and are therefore relevant to normal mRNA turnover. Third, a truncated form of eIF4E (Delta196) had a reduced capacity to inhibit Dcp1-mediated decapping in vitro, yet did not change cellular mRNA half-lives. Thus, the accessibility of the cap to Dcp1 in vivo is not simply controlled by competition with eIF4E, but is subject to switching between molecular states with different levels of access.
在酿酒酵母中,由Dcp1介导的去帽作用是mRNA降解的关键步骤。然而,帽结构还会与真核生物起始因子(eIF)复合物4F及其相关蛋白结合。表征去帽作用与涉及eIF4F的相互作用之间的关系,是理解多核糖体解体和mRNA衰变的重要一步。三类观察结果表明了eIF4F功能状态的变化如何在体内调节mRNA的稳定性。首先,由eIF4E突变或酵母4E结合蛋白p20的存在导致的eIF4E与eIF4G之间相互作用的部分破坏,使mRNA得以稳定。因此,eIF4G和p20与eIF4E的相互作用可能起到调节去帽过程的作用。由于我们还表明体外去帽速率不受mRNA主体性质的直接影响,这表明eIF4F结构的变化可能在mRNA衰变过程中触发去帽作用。其次,这些效应在细胞整体翻译速率没有极端变化的情况下也能观察到,因此与正常的mRNA周转相关。第三,截短形式的eIF4E(Delta196)在体外抑制Dcp1介导的去帽作用的能力降低,但并未改变细胞内mRNA的半衰期。因此,帽结构在体内对Dcp1的可及性并非简单地由与eIF4E的竞争所控制,而是会在具有不同可及水平的分子状态之间切换。