Delaforge Elise, Due Amanda D, Theisen Frederik Friis, Morffy Nicolas, O'Shea Charlotte, Blackledge Martin, Strader Lucia C, Skriver Karen, Kragelund Birthe B
REPIN, Department of Biology, University of Copenhagen, Ole Maaløes vej 5, DK-2200 Copenhagen N, Denmark.
Linderstrøm-Lang Centre for Protein Science and Department of Biology, University of Copenhagen, Ole Maaløes vej 5, DK-2200 Copenhagen N, Denmark.
Nucleic Acids Res. 2025 Feb 8;53(4). doi: 10.1093/nar/gkaf065.
Transcriptional regulation involves interactions between transcription factors, coregulators, and DNA. Intrinsic disorder is a major player in this regulation, but mechanisms driven by disorder remain elusive. Here, we address molecular communication within the stress-regulating Arabidopsis thaliana transcription factor ANAC013. Through high-throughput screening of ANAC013 for transcriptional activation activity, we identify three activation domains within its C-terminal intrinsically disordered region. Two of these overlap with acidic islands and form dynamic interactions with the DNA-binding domain and are released, not only upon binding of target promoter DNA, but also by nonspecific DNA. We show that independently of DNA binding, the RST (RCD--SRO--TAF4) domain of the negative regulator RCD1 (Radical-induced Cell Death1) scavenges the two acidic activation domains positioned vis-à-vis through allovalent binding, leading to dynamic occupation at enhanced affinity. We propose an allovalency model for transcriptional regulation, where sequentially close activation domains in both DNA-bound and DNA-free states allow for efficient regulation. The model is likely relevant for many transcription factor systems, explaining the functional advantage of carrying sequentially close activation domains.
转录调控涉及转录因子、共调节因子和DNA之间的相互作用。内在无序是这种调控的主要参与者,但由无序驱动的机制仍然难以捉摸。在这里,我们研究了应激调节拟南芥转录因子ANAC013内的分子通讯。通过对ANAC013进行转录激活活性的高通量筛选,我们在其C端内在无序区域内鉴定出三个激活域。其中两个与酸性岛重叠,并与DNA结合域形成动态相互作用,不仅在结合靶启动子DNA时释放,而且也可被非特异性DNA释放。我们表明,负调节因子RCD1(自由基诱导细胞死亡1)的RST(RCD - SRO - TAF4)结构域独立于DNA结合,通过异价结合清除相对定位的两个酸性激活域,导致以增强的亲和力进行动态占据。我们提出了一种转录调控的异价模型,其中在DNA结合和非DNA结合状态下顺序紧密的激活域允许进行有效调控。该模型可能与许多转录因子系统相关,解释了携带顺序紧密的激活域的功能优势。