Key Laboratory of the Ministry of Education for Coastal and Wetland Ecosystems, College of the Environment and Ecology, Xiamen University, Xiamen, Fujian, China 361102.
Graduate College of Biomedical Sciences, Western University of Health Sciences, Pomona, California 91766.
Plant Cell. 2019 Oct;31(10):2332-2352. doi: 10.1105/tpc.18.00545. Epub 2019 Aug 19.
A crucial step for mRNA polyadenylation is poly(A) signal recognition by -acting factors. The mammalian cleavage and polyadenylation specificity factor (CPSF) complex components CPSF30 and WD repeat-containing protein33 (WDR33) recognize the canonical AAUAAA for polyadenylation. In Arabidopsis (), the flowering time regulator FY is the homolog of WDR33. However, its role in mRNA polyadenylation is poorly understood. Using poly(A) tag sequencing, we found that >50% of alternative polyadenylation (APA) events are altered in single mutants or double mutants with (a null mutant of ), but mutation of the FY WD40-repeat has a stronger effect than deletion of the plant-unique Pro-Pro-Leu-Pro-Pro (PPLPP) domain. mutations disrupt AAUAAA or AAUAAA-like poly(A) signal recognition. Notably, A-rich signal usage is suppressed in the WD40-repeat mutation but promoted in PPLPP-domain deficiency. However, mutations do not aggravate the altered signal usage in Furthermore, the WD40-repeat mutation shows a preference for 3' untranslated region shortening, but the PPLPP-domain deficiency shows a preference for lengthening. Interestingly, the WD40-repeat mutant exhibits shortened primary roots and late flowering with alteration of APA of related genes. Importantly, the long transcripts of two APA genes affected in are related to abiotic stress responses. These results reveal a conserved and specific role of FY in mRNA polyadenylation.
聚腺苷酸化的关键步骤是-作用因子识别聚(A)信号。哺乳动物切割多聚腺苷酸化特异性因子(CPSF)复合物的组成部分 CPSF30 和 WD 重复蛋白 33(WDR33)识别聚腺苷酸化的典型 AAUAAA。在拟南芥()中,开花时间调节剂 FY 是 WDR33 的同源物。然而,其在 mRNA 聚腺苷酸化中的作用知之甚少。通过聚(A)标签测序,我们发现 FY 单突变体或与 FY(一个 FY 的 null 突变体)双突变体中的 >50%的可变聚腺苷酸化(APA)事件发生改变,但 FY WD40 重复突变的影响比植物特有的 Pro-Pro-Leu-Pro-Pro(PPLPP)结构域缺失更强。FY 突变破坏 AAUAAA 或 AAUAAA 样聚(A)信号识别。值得注意的是,在 WD40 重复突变中,富含 A 的信号使用受到抑制,但在 PPLPP 结构域缺失中受到促进。然而,FY 突变并没有加剧 FY 缺失中改变的信号使用。此外,WD40 重复突变表现出 3'非翻译区缩短的偏好,而 PPLPP 结构域缺失则表现出延长的偏好。有趣的是,WD40 重复突变体表现出缩短的主根和开花较晚,同时 APA 相关基因发生改变。重要的是,FY 缺失中受影响的两个 APA 基因的长转录本与非生物胁迫反应有关。这些结果揭示了 FY 在 mRNA 聚腺苷酸化中保守而特异的作用。