Paul Biplab, Montpetit Ben
Department of Cell Biology, University of Alberta, Edmonton, AB T6G 2H7, Canada.
Department of Cell Biology, University of Alberta, Edmonton, AB T6G 2H7, Canada
Mol Biol Cell. 2016 Sep 1;27(17):2742-56. doi: 10.1091/mbc.E16-04-0244. Epub 2016 Jul 6.
Many protein factors are required for mRNA biogenesis and nuclear export, which are central to the eukaryotic gene expression program. It is unclear, however, whether all factors have been identified. Here we report on a screen of >1000 essential gene mutants in Saccharomyces cerevisiae for defects in mRNA processing and export, identifying 26 mutants with defects in this process. Single-molecule FISH data showed that the majority of these mutants accumulated mRNA within specific regions of the nucleus, which included 1) mRNAs within the nucleolus when nucleocytoplasmic transport, rRNA biogenesis, or RNA processing and surveillance was disrupted, 2) the buildup of mRNAs near transcription sites in 3'-end processing and chromosome segregation mutants, and 3) transcripts being enriched near nuclear pore complexes when components of the mRNA export machinery were mutated. These data show that alterations to various nuclear processes lead to the retention of mRNAs at discrete locations within the nucleus.
mRNA生物合成和核输出需要许多蛋白质因子,这些过程是真核基因表达程序的核心。然而,目前尚不清楚是否已鉴定出所有因子。在此,我们报告了一项针对酿酒酵母中1000多个必需基因突变体的筛选,以检测mRNA加工和输出缺陷,鉴定出26个在此过程中存在缺陷的突变体。单分子荧光原位杂交数据显示,这些突变体中的大多数在细胞核的特定区域积累mRNA,其中包括:1)当核质运输、rRNA生物合成或RNA加工及监测被破坏时,核仁内的mRNA;2)在3'端加工和染色体分离突变体中,mRNA在转录位点附近积累;3)当mRNA输出机制的组分发生突变时,转录本在核孔复合体附近富集。这些数据表明,各种核过程的改变会导致mRNA在细胞核内的离散位置滞留。