Instituto de Biología Molecular y Celular de Plantas (CSIC-UPV), 46022 Valencia, Spain.
Ecole Normale Supérieure, Institut de Biologie de l'Ecole Normale Supérieure (CNRS), CNRS, INSERM, Université PSL, 75230 Paris, France.
Plant Physiol. 2024 Apr 30;195(1):640-651. doi: 10.1093/plphys/kiae041.
The evolutionarily conserved POLYMERASE-ASSOCIATED FACTOR1 complex (Paf1C) participates in transcription, and research in animals and fungi suggests that it facilitates RNA POLYMERASE II (RNAPII) progression through chromatin. We examined the genomic distribution of the EARLY FLOWERING7 (ELF7) and VERNALIZATION INDEPENDENCE3 subunits of Paf1C in Arabidopsis (Arabidopsis thaliana). The occupancy of both subunits was confined to thousands of gene bodies and positively associated with RNAPII occupancy and the level of gene expression, supporting a role as a transcription elongation factor. We found that monoubiquitinated histone H2B, which marks most transcribed genes, was strongly reduced genome wide in elf7 seedlings. Genome-wide profiling of RNAPII revealed that in elf7 mutants, RNAPII occupancy was reduced throughout the gene body and at the transcription end site of Paf1C-targeted genes, suggesting a direct role for the complex in transcription elongation. Overall, our observations suggest a direct functional link between Paf1C activity, monoubiquitination of histone H2B, and the transition of RNAPII to productive elongation. However, for several genes, Paf1C may also act independently of H2Bub deposition or occupy these genes more stable than H2Bub marking, possibly reflecting the dynamic nature of Paf1C association and H2Bub turnover during transcription.
进化保守的 POLYMERASE-ASSOCIATED FACTOR1 复合物(Paf1C)参与转录,动物和真菌的研究表明,它有助于 RNA POLYMERASE II(RNAPII)在染色质中前进。我们检查了 EARLY FLOWERING7(ELF7)和 Paf1C 的 VERNALIZATION INDEPENDENCE3 亚基在拟南芥(Arabidopsis thaliana)中的基因组分布。这两个亚基的占据都局限于数千个基因体,与 RNAPII 的占据和基因表达水平呈正相关,支持作为转录延伸因子的作用。我们发现,单泛素化组蛋白 H2B,标记大多数转录基因,在 elf7 幼苗中广泛减少。RNAPII 的全基因组分析表明,在 elf7 突变体中,RNAPII 占据在基因体和 Paf1C 靶向基因的转录末端位点都减少了,这表明该复合物在转录延伸中具有直接作用。总的来说,我们的观察结果表明,Paf1C 活性、组蛋白 H2B 的单泛素化以及 RNAPII 向生产性延伸的转变之间存在直接的功能联系。然而,对于几个基因,Paf1C 可能也独立于 H2Bub 沉积起作用,或者比 H2Bub 标记更稳定地占据这些基因,这可能反映了 Paf1C 关联和 H2Bub 周转在转录过程中的动态性质。