Bhat Prashant, Chow Amy, Emert Benjamin, Ettlin Olivia, Quinodoz Sofia A, Takei Yodai, Huang Wesley, Blanco Mario R, Guttman Mitchell
bioRxiv. 2023 Jan 4:2023.01.04.522632. doi: 10.1101/2023.01.04.522632.
The nucleus is highly organized such that factors involved in transcription and processing of distinct classes of RNA are organized within specific nuclear bodies. One such nuclear body is the nuclear speckle, which is defined by high concentrations of protein and non-coding RNA regulators of pre-mRNA splicing. What functional role, if any, speckles might play in the process of mRNA splicing remains unknown. Here we show that genes localized near nuclear speckles display higher spliceosome concentrations, increased spliceosome binding to their pre-mRNAs, and higher co-transcriptional splicing levels relative to genes that are located farther from nuclear speckles. We show that directed recruitment of a pre-mRNA to nuclear speckles is sufficient to drive increased mRNA splicing levels. Finally, we show that gene organization around nuclear speckles is highly dynamic with differential localization between cell types corresponding to differences in Pol II occupancy. Together, our results integrate the longstanding observations of nuclear speckles with the biochemistry of mRNA splicing and demonstrate a critical role for dynamic 3D spatial organization of genomic DNA in driving spliceosome concentrations and controlling the efficiency of mRNA splicing.
细胞核高度有序,使得参与不同类型RNA转录和加工的因子在特定的核体内有序排列。其中一种核体是核斑,它由高浓度的前体mRNA剪接的蛋白质和非编码RNA调节因子所界定。核斑在mRNA剪接过程中可能发挥何种功能作用(如果有的话)仍然未知。在这里,我们表明,相对于远离核斑的基因,定位在核斑附近的基因显示出更高的剪接体浓度、剪接体与其前体mRNA的结合增加以及更高的共转录剪接水平。我们表明,将前体mRNA定向募集到核斑足以驱动mRNA剪接水平的提高。最后,我们表明,核斑周围的基因组织具有高度动态性,细胞类型之间的差异定位对应于RNA聚合酶II占据率的差异。总之,我们的结果将对核斑的长期观察与mRNA剪接的生物化学相结合,并证明基因组DNA的动态三维空间组织在驱动剪接体浓度和控制mRNA剪接效率方面起着关键作用。