{"title":"拟南芥转录因子 LATE FLOWERING 的基因组编辑和分子分析。","authors":"Yoshimi Nakano, Maki Kawai, Moeca Arai, Sumire Fujiwara","doi":"10.5511/plantbiotechnology.23.0920a","DOIUrl":null,"url":null,"abstract":"<p><p>Correct flower organ formation at the right timing is one of the most important strategies for plants to achieve reproductive success. Ectopic overexpression of LATE FLOWERING (LATE) is known to induce late flowering, partly through suppressing expression of the florigen-encoding gene <i>FLOWERING LOCUS T</i> (<i>FT</i>) in <i>Arabidopsis</i>. LATE is one of the C2H2 zinc finger transcription factors, and it has a canonical transcriptional repression domain called the ethylene-responsive element-binding factor-associated amphiphilic repression (EAR) motif at the end of its C terminus. Therefore, LATE is considered a transcriptional repressor, but its molecular function remains unclear. Our genome-edited <i>late</i> mutants exhibited no distinct phenotype, even in flowering, indicating the presence of redundancy from other factors. To reveal the molecular function of LATE and factors working with it, we investigated its transcriptional activity and interactions with other proteins. Transactivation activity assay showed that LATE possesses transcriptional repression ability, which appears to be attributable to both the EAR motif and other sequences. Yeast two-hybrid assay showed the EAR motif-mediated interaction of LATE with TOPLESS, a transcriptional corepressor. Moreover, LATE could also interact with CRABS CLAW (CRC), one of the most important regulators of floral meristem determinacy, through sequences in LATE other than the EAR motif. 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引用次数: 0
摘要
在正确的时间形成正确的花器官是植物获得繁殖成功的最重要策略之一。在拟南芥中,异位过表达晚开花(LATE)可诱导晚开花,部分原因是它抑制了花原编码基因FLOWERING LOCUS T(FT)的表达。LATE 是 C2H2 锌指转录因子之一,它的 C 端末端有一个典型的转录抑制结构域,称为乙烯反应元件结合因子相关两性抑制(EAR)结构域。因此,LATE 被认为是一种转录抑制因子,但其分子功能仍不清楚。我们的基因组编辑的晚期突变体没有表现出明显的表型,即使在开花期也是如此,这表明存在其他因子的冗余。为了揭示 LATE 及其作用因子的分子功能,我们研究了它的转录活性以及与其他蛋白的相互作用。转录激活活性分析表明,LATE具有转录抑制能力,这似乎可归因于EAR基序和其他序列。酵母双杂交实验表明,LATE与转录核心抑制因子TOPLESS之间存在EAR基序介导的相互作用。此外,LATE 还能通过 EAR 基序以外的序列与花分生组织决定性的最重要调控因子之一 CRABS CLAW(CRC)相互作用。我们的研究结果表明,LATE 有可能与 CRC 形成转录抑制复合物,以实现花分生组织的决定性。
Genome editing and molecular analyses of an Arabidopsis transcription factor, LATE FLOWERING.
Correct flower organ formation at the right timing is one of the most important strategies for plants to achieve reproductive success. Ectopic overexpression of LATE FLOWERING (LATE) is known to induce late flowering, partly through suppressing expression of the florigen-encoding gene FLOWERING LOCUS T (FT) in Arabidopsis. LATE is one of the C2H2 zinc finger transcription factors, and it has a canonical transcriptional repression domain called the ethylene-responsive element-binding factor-associated amphiphilic repression (EAR) motif at the end of its C terminus. Therefore, LATE is considered a transcriptional repressor, but its molecular function remains unclear. Our genome-edited late mutants exhibited no distinct phenotype, even in flowering, indicating the presence of redundancy from other factors. To reveal the molecular function of LATE and factors working with it, we investigated its transcriptional activity and interactions with other proteins. Transactivation activity assay showed that LATE possesses transcriptional repression ability, which appears to be attributable to both the EAR motif and other sequences. Yeast two-hybrid assay showed the EAR motif-mediated interaction of LATE with TOPLESS, a transcriptional corepressor. Moreover, LATE could also interact with CRABS CLAW (CRC), one of the most important regulators of floral meristem determinacy, through sequences in LATE other than the EAR motif. Our findings demonstrated the possibility that LATE can form a transcriptional repression complex with CRC for floral meristem determinacy.
期刊介绍:
Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance.
Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.