A single helix repression domain is functional across diverse eukaryotes.
A single helix repression domain is functional across diverse eukaryotes.
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DOI：
10.1073/pnas.2206986119
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发表时间：
2022-10-11
影响因子：
11.1
通讯作者：
中科院分区：
文献类型：
--
作者：
Lis 1同源（LisH）结构域存在于近25，000种具有不同功能的蛋白质中，包括几种辅阻遏物。以前，我们发现，在植物辅阻遏物的LisH结构域的第一个螺旋（H1）作为一个短（18个氨基酸），模块化的阻遏结构域。在这里，我们通过调查代表真菌，动物和植物的1,000多种蛋白质的LisH-H1的功能来扩展这种分析。大多数测试序列抑制酵母中的报告活性，并且当在植物中测试时，许多保留了这种功能，这表明抑制是LisH结构域的进化保守作用。我们进一步调整了我们的测定以研究对两种含有LisH结构域的人类蛋白质TBL 1和DCAF 1中癌症相关突变的抑制活性的影响。辅阻遏物TOPLESS（TPL）及其旁系同源物协同调节植物发育和免疫的大量关键基因。在更大的泛真核Tup 1/TLE/Groucho辅阻遏蛋白家族的许多成员中，TPL包含一个Lis 1同源结构域（LisH），其功能还不清楚。我们先前已经发现TPL中的LisH，特别是N-末端18个氨基酸的α-螺旋区（TPL-H1），可以作为一个自主阻遏结构域。我们假设，不同的LisH蛋白的同源结构域可以共享相同的功能。为了验证这一假设，我们建立了一个H1 s文库，广泛采样LisH结构域的序列和进化空间，并在酿酒酵母的合成转录抑制试验中测试了它们的活性。使用这种方法，我们发现抑制活性是高度保守的，并且可能是该基序的祖先功能。我们还确定了有助于抑制功能的关键残基。我们将这一新知识用于两个应用。首先，我们测试了体细胞癌症中发现的突变对两种含LisH的人类蛋白质的抑制功能的作用。其次，我们验证了我们的许多阻遏结构域在植物中的功能，证实了这些序列应该用于许多真核生物的合成生物学应用。
The Lis1 Homology (LisH) domain is found in nearly 25,000 proteins with diverse functions, including several corepressors. Previously, we found that the first helix (H1) of the LisH domain in a plant corepressor functioned as a short (18 amino acids), modular repression domain. Here, we extended this analysis by surveying the function of LisH-H1s representing over 1,000 proteins from fungi, animals, and plants. The majority of the tested sequences repressed reporter activity in yeast, and many retained this function when tested in plants, pointing to repression as an evolutionarily conserved role for LisH domains. We further adapted our assay to study the impact on repressive activity of cancer-associated mutations in two LisH-domain containing human proteins: TBL1 and DCAF1. The corepressor TOPLESS (TPL) and its paralogs coordinately regulate a large number of genes critical to plant development and immunity. As in many members of the larger pan-eukaryotic Tup1/TLE/Groucho corepressor family, TPL contains a Lis1 Homology domain (LisH), whose function is not well understood. We have previously found that the LisH in TPL—and specifically the N-terminal 18 amino acid alpha-helical region (TPL-H1)—can act as an autonomous repression domain. We hypothesized that homologous domains across diverse LisH-containing proteins could share the same function. To test that hypothesis, we built a library of H1s that broadly sampled the sequence and evolutionary space of LisH domains, and tested their activity in a synthetic transcriptional repression assay in Saccharomyces cerevisiae. Using this approach, we found that repression activity was highly conserved and likely the ancestral function of this motif. We also identified key residues that contribute to repressive function. We leveraged this new knowledge for two applications. First, we tested the role of mutations found in somatic cancers on repression function in two human LisH-containing proteins. Second, we validated function of many of our repression domains in plants, confirming that these sequences should be of use to synthetic biology applications across many eukaryotes.
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