Instituto de Biomedicina de Sevilla, Universidad de Sevilla-CSIC-Hospital Universitario V. Del Rocío, Seville, Spain.
Departamento de Genética, Facultad de Biología, Universidad de Sevilla, Seville, Spain.
RNA Biol. 2021 Sep;18(9):1310-1323. doi: 10.1080/15476286.2020.1845504. Epub 2020 Dec 1.
mRNA homoeostasis is favoured by crosstalk between transcription and degradation machineries. Both the Ccr4-Not and the Xrn1-decaysome complexes have been described to influence transcription. While Ccr4-Not has been shown to directly stimulate transcription elongation, the information available on how Xrn1 influences transcription is scarce and contradictory. In this study we have addressed this issue by mapping RNA polymerase II (RNA pol II) at high resolution, using CRAC and BioGRO-seq techniques in . We found significant effects of Xrn1 perturbation on RNA pol II profiles across the genome. RNA pol II profiles at 5' exhibited significant alterations that were compatible with decreased elongation rates in the absence of Xrn1. Nucleosome mapping detected altered chromatin configuration in the gene bodies. We also detected accumulation of RNA pol II shortly upstream of polyadenylation sites by CRAC, although not by BioGRO-seq, suggesting higher frequency of backtracking before pre-mRNA cleavage. This phenomenon was particularly linked to genes with poorly positioned nucleosomes at this position. Accumulation of RNA pol II at 3' was also detected in other mRNA decay mutants. According to these and other pieces of evidence, Xrn1 seems to influence transcription elongation at least in two ways: by directly favouring elongation rates and by a more general mechanism that connects mRNA decay to late elongation.
mRNA 动态平衡受到转录和降解机制之间相互作用的影响。Ccr4-Not 和 Xrn1 衰变体复合物都被描述为影响转录。虽然 Ccr4-Not 被证明可以直接刺激转录延伸,但关于 Xrn1 如何影响转录的信息很少且相互矛盾。在这项研究中,我们通过使用 CRAC 和 BioGRO-seq 技术在. 中高分辨率地映射 RNA 聚合酶 II (RNA pol II) 来解决这个问题。我们发现 Xrn1 扰动对全基因组中 RNA pol II 图谱有显著影响。5'端的 RNA pol II 图谱显示出明显的变化,与 Xrn1 缺失时的延伸率降低相兼容。核小体作图检测到基因体内染色质构型的改变。我们还通过 CRAC 检测到在 polyadenylation 位点上游 RNA pol II 的短暂积累,但通过 BioGRO-seq 未检测到,这表明在 pre-mRNA 切割之前,回溯的频率更高。这种现象与在该位置处位置不佳的核小体的基因特别相关。在其他 mRNA 衰变突变体中也检测到 RNA pol II 在 3'端的积累。根据这些和其他证据,Xrn1 似乎至少以两种方式影响转录延伸:直接促进延伸率,以及一种将 mRNA 衰变与晚期延伸联系起来的更普遍机制。