Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.
University of Chinese Academy of Sciences, Beijing 100049, China.
Plant Physiol. 2019 Feb;179(2):686-699. doi: 10.1104/pp.18.00782. Epub 2018 Nov 28.
Polyadenylation influences gene expression by affecting mRNA stability, transport, and translatability. Here, we report that Cleavage stimulation Factor 77 (AtCstF77), a component of the pre-mRNA 3'-end polyadenylation machinery, affects polyadenylation site (PAS) selection in transcripts of some auxin signaling genes in Arabidopsis (). Disruption of reduced auxin sensitivity and decreased the expression of the auxin reporter - Null mutations of caused severe developmental defects, but were not lethal as previously reported. - genetically interacted with - auxin receptor double mutants, further supporting that polyadenylation affects auxin signaling. was ubiquitously expressed in embryos, seedlings, and adult plants. The AtCstF77 protein was localized in the nucleus, which is consistent with its function in pre-mRNA processing. We observed that PASs in transcripts from approximately 2,400 genes were shifted in the - mutant. Moreover, most of the PAS shifts were from proximal to distal sites. Auxin treatment also caused PAS shifts in transcripts from a small number of genes. Several auxin signaling or homeostasis genes had different PASs in their transcripts in the - mutant. The expression levels of /-- were significantly increased in the - mutant, which can partially account for the auxin resistance phenotype of this mutant. Our results demonstrate that AtCstF77 plays pleiotropic and critical roles in Arabidopsis development. Moreover, disruption of AtCstF64, another component of the polyadenylation machinery, led to developmental defects and reduced auxin response, similar to those of the - mutant. We conclude that AtCstF77 affects auxin responses, likely by controlling PAS selection of transcripts of some auxin signaling components.
多聚腺苷酸化通过影响 mRNA 的稳定性、运输和可翻译性来影响基因表达。在这里,我们报告说,Cleavage stimulation Factor 77(AtCstF77),一种前体 mRNA 3'-端多聚腺苷酸化机制的组成部分,影响拟南芥中一些生长素信号基因转录物的多聚腺苷酸化位点(PAS)选择()。 破坏 减少了生长素敏感性并降低了生长素报告者的表达 - 的双突变体与生长素受体的缺失突变体相互作用,进一步支持多聚腺苷酸化影响生长素信号转导。 在胚胎、幼苗和成年植物中广泛表达。AtCstF77 蛋白定位于细胞核中,这与其在 pre-mRNA 加工中的功能一致。我们观察到在 - 突变体中,约 2400 个基因的转录物中的 PAS 发生了移位。此外,大多数 PAS 移位是从近端到远端位点。生长素处理也会导致少数基因转录物中的 PAS 发生移位。一些生长素信号或稳态基因在其 - 突变体的转录物中具有不同的 PAS。/--的表达水平在 - 突变体中显著增加,这可以部分解释该突变体的生长素抗性表型。我们的结果表明,AtCstF77 在拟南芥发育中发挥着多种关键作用。此外,另一种多聚腺苷酸化机制的组成部分 AtCstF64 的破坏导致了发育缺陷和生长素反应降低,与 - 突变体的表型相似。我们得出结论,AtCstF77 影响生长素反应,可能是通过控制一些生长素信号成分的转录物的 PAS 选择。