Cell Biology & Plant Biochemistry, Biochemistry Centre, University of Regensburg, Universitätsstr. 31, D-93053 Regensburg, Germany.
NGS Analysis Centre, Biology and Pre-Clinical Medicine, University of Regensburg, Universitätsstr. 31, D-93053 Regensburg, Germany.
Nucleic Acids Res. 2023 Nov 27;51(21):11518-11533. doi: 10.1093/nar/gkad825.
Various transcript elongation factors (TEFs) including modulators of RNA polymerase II (RNAPII) activity and histone chaperones tune the efficiency of transcription in the chromatin context. TEFs are involved in establishing gene expression patterns during growth and development in Arabidopsis, while little is known about the genomic distribution of the TEFs and the way they facilitate transcription. We have mapped the genome-wide occupancy of the elongation factors SPT4-SPT5, PAF1C and FACT, relative to that of elongating RNAPII phosphorylated at residues S2/S5 within the carboxyterminal domain. The distribution of SPT4-SPT5 along transcribed regions closely resembles that of RNAPII-S2P, while the occupancy of FACT and PAF1C is rather related to that of RNAPII-S5P. Under transcriptionally challenging heat stress conditions, mutant plants lacking the corresponding TEFs are differentially impaired in transcript synthesis. Strikingly, in plants deficient in PAF1C, defects in transcription across intron/exon borders are observed that are cumulative along transcribed regions. Upstream of transcriptional start sites, the presence of FACT correlates with nucleosomal occupancy. Under stress conditions FACT is particularly required for transcriptional upregulation and to promote RNAPII transcription through +1 nucleosomes. Thus, Arabidopsis TEFs are differently distributed along transcribed regions, and are distinctly required during transcript elongation especially upon transcriptional reprogramming.
各种转录延伸因子(TEFs),包括 RNA 聚合酶 II(RNAPII)活性调节剂和组蛋白伴侣,在染色质环境中调节转录效率。TEFs 参与了拟南芥生长和发育过程中基因表达模式的建立,而关于 TEFs 的基因组分布以及它们促进转录的方式知之甚少。我们已经绘制了延伸因子 SPT4-SPT5、PAF1C 和 FACT 在全基因组范围内相对于羧基末端结构域中磷酸化的延伸 RNAPII 的占有率。SPT4-SPT5 在转录区域的分布与 RNAPII-S2P 非常相似,而 FACT 和 PAF1C 的占有率与 RNAPII-S5P 相当。在转录挑战性的热应激条件下,缺乏相应 TEFs 的突变体植物在转录物合成中受到不同程度的损伤。引人注目的是,在 PAF1C 缺陷型植物中,观察到跨内含子/外显子边界的转录缺陷,这些缺陷在转录区域中累积。在转录起始位点上游,FACT 的存在与核小体占有率相关。在应激条件下,FACT 特别需要促进转录上调,并通过+1 核小体促进 RNAPII 转录。因此,拟南芥 TEFs 在转录区域中呈不同分布,在转录延伸过程中,特别是在转录重编程时,具有明显的需求。