Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Hongo, Tokyo, Japan.
EMBO J. 2010 Jul 7;29(13):2173-81. doi: 10.1038/emboj.2010.108. Epub 2010 May 28.
A number of meiosis-specific mRNAs are initially weakly transcribed, but then selectively removed during fission yeast mitotic growth. These mRNAs harbour a region termed DSR (determinant of selective removal), which is recognized by the YTH family RNA-binding protein Mmi1p. Mmi1p directs the destruction of these mRNAs in collaboration with nuclear exosomes. However, detailed molecular mechanisms underlying this process of selective mRNA elimination have remained elusive. In this study, we demonstrate the critical role of polyadenylation in this process. Two-hybrid and genetic screens revealed potential interactions between Mmi1p and proteins involved in polyadenylation. Additional investigations showed that destruction of DSR-containing mRNAs by exosomes required polyadenylation by a canonical poly(A) polymerase. The recruitment of Pab2p, a poly(A)-binding protein, to the poly(A) tail was also necessary for mRNA destruction. In cells undergoing vegetative growth, Mmi1p localized with exosomes, Pab2p, and components of the polyadenylation complex in several patchy structures in the nucleoplasm. These patches may represent the sites for degradation of meiosis-specific mRNAs with untimely expression.
许多减数分裂特异性 mRNAs 最初被弱转录,但随后在裂殖酵母有丝分裂生长过程中被选择性去除。这些 mRNAs 含有一个称为 DSR(选择性去除决定因素)的区域,该区域被 YTH 家族 RNA 结合蛋白 Mmi1p 识别。Mmi1p 与核外切体协同指导这些 mRNAs 的破坏。然而,这个选择性 mRNA 消除过程的详细分子机制仍然难以捉摸。在这项研究中,我们证明了多聚腺苷酸化在这个过程中的关键作用。双杂交和遗传筛选揭示了 Mmi1p 与参与多聚腺苷酸化的蛋白质之间的潜在相互作用。进一步的研究表明,外切体对含有 DSR 的 mRNAs 的破坏需要由一个规范的 poly(A) 聚合酶进行多聚腺苷酸化。Pab2p,一种 poly(A)-结合蛋白,被招募到 poly(A) 尾巴也是 mRNA 破坏所必需的。在进行营养生长的细胞中,Mmi1p 与外切体、Pab2p 和多聚腺苷酸化复合物的成分一起定位于核质中的几个斑片状结构中。这些斑点可能代表有丝分裂特异性 mRNAs 表达过早时降解的部位。