Department of Biology, University of Washington, Seattle, WA, 98195-1800, USA.
Department of Horticulture, Nanjing Agricultural University, Nanjing, 210095, China.
Plant J. 2017 Oct;92(2):244-262. doi: 10.1111/tpj.13649. Epub 2017 Sep 5.
CYCLING DOF FACTOR 1 (CDF1) and its homologs play an important role in the floral transition by repressing the expression of floral activator genes such as CONSTANS (CO) and FLOWERING LOCUS T (FT) in Arabidopsis. The day-length-specific removal of CDF1-dependent repression is a critical mechanism in photoperiodic flowering. However, the mechanism by which CDF1 represses CO and FT transcription remained elusive. Here we demonstrate that Arabidopsis CDF proteins contain non-EAR motif-like conserved domains required for interaction with the TOPLESS (TPL) co-repressor protein. This TPL interaction confers a repressive function on CDF1, as mutations of the N-terminal TPL binding domain largely impair the ability of CDF1 protein to repress its targets. TPL proteins are present on specific regions of the CO and FT promoters where CDF1 binds during the morning. In addition, TPL binding increases when CDF1 expression is elevated, suggesting that TPL is recruited to these promoters in a time-dependent fashion by CDFs. Moreover, reduction of TPL activity induced by expressing a dominant negative version of TPL (tpl-1) in phloem companion cells results in early flowering and a decreased sensitivity to photoperiod in a manner similar to a cdf loss-of-function mutant. Our results indicate that the mechanism of CDF1 repression is through the formation of a CDF-TPL transcriptional complex, which reduces the expression levels of CO and FT during the morning for seasonal flowering.
循环 DOF 因子 1(CDF1)及其同源物在花发育转变中发挥重要作用,通过抑制拟南芥中花发育促进因子基因如 CONSTANS(CO)和 FLOWERING LOCUS T(FT)的表达。CDF1 依赖性抑制作用的日长特异性去除是光周期开花的一个关键机制。然而,CDF1 抑制 CO 和 FT 转录的机制仍不清楚。在这里,我们证明拟南芥 CDF 蛋白含有非 EAR 基序样保守结构域,这些结构域对于与 TOPLESS(TPL)共抑制蛋白相互作用是必需的。这种 TPL 相互作用赋予 CDF1 抑制功能,因为 N 端 TPL 结合结构域的突变在很大程度上削弱了 CDF1 蛋白抑制其靶标的能力。TPL 蛋白存在于 CO 和 FT 启动子的特定区域,CDF1 在早晨结合这些区域。此外,当 CDF1 表达升高时,TPL 结合增加,这表明 TPL 以时间依赖性方式被 CDFs 招募到这些启动子。此外,通过在韧皮部伴胞中表达显性负突变体 TPL(tpl-1)来降低 TPL 活性,导致开花提前并且对光周期的敏感性降低,这与 cdf 功能丧失突变体的表型相似。我们的结果表明,CDF1 抑制的机制是通过形成 CDF-TPL 转录复合物,该复合物在早晨降低 CO 和 FT 的表达水平,从而促进季节性开花。