The TRIM33 Bromodomain Recognizes Histone Lysine Lactylation.

IF 3.5 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Raymundo Nuñez, Paul F W Sidlowski, Erica A Steen, Sarah L Wynia-Smith, Daniel J Sprague, Robert F Keyes, Brian C Smith
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引用次数: 0

Abstract

Histone lysine lactylation (Kla) regulates inflammatory gene expression in activated macrophages and mediates the polarization of inflammatory (M1) to reparative (M2) macrophages. However, the molecular mechanisms and key protein players involved in Kla-mediated transcriptional changes are unknown. As Kla is structurally similar to lysine acetylation (Kac), which is bound by bromodomains, we hypothesized that bromodomain-containing proteins bind histone Kla. Here, we screened 28 recombinantly expressed bromodomains for binding to histone Kla peptides via AlphaScreen assays. TRIM33 was the sole bromodomain tested that bound histone Kla peptides. TRIM33 attenuates inflammatory genes during late-stage macrophage activation; thus, TRIM33 provides a potential link between histone Kla and macrophage polarization. Orthogonal biophysical techniques, including isothermal titration calorimetry and protein-detected nuclear magnetic resonance, confirmed the submicromolar binding affinity of the TRIM33 bromodomain to both Kla and Kac histone post-translational modifications. Sequence alignments of human bromodomains revealed a unique glutamic acid residue within the TRIM33 binding pocket that we found confers TRIM33 specificity for binding Kla compared with other bromodomains. Molecular modeling of interactions of Kla with the TRIM33 bromodomain binding pocket and site-directed mutagenesis of glutamic acid confirmed the critical role of this residue in the selective recognition of Kla by TRIM33. Collectively, our findings implicate TRIM33, a bromodomain-containing protein, as a novel reader of histone Kla, potentially bridging the gap between histone Kla and macrophage polarization. This study enhances our understanding of the regulatory role of histone Kla in macrophage-mediated inflammation and offers insights into the underlying structural and biophysical mechanisms.

TRIM33 溴odomain 可识别组蛋白赖氨酸乳化。
组蛋白赖氨酸乳酰化(Kla)调节活化巨噬细胞中炎症基因的表达,并介导炎症(M1)巨噬细胞向修复(M2)巨噬细胞的极化。然而,参与 Kla 介导的转录变化的分子机制和关键蛋白参与者尚不清楚。由于 Kla 在结构上类似于赖氨酸乙酰化(Kac),而赖氨酸乙酰化(Kac)是由溴基团结合的,因此我们推测含溴基团的蛋白会结合组蛋白 Kla。在这里,我们通过 AlphaScreen 分析筛选了 28 个重组表达的溴化多聚酶链与组蛋白 Kla 肽的结合。TRIM33 是所测试的唯一能与组蛋白 Kla 多肽结合的溴结构域。在巨噬细胞活化晚期,TRIM33可抑制炎症基因;因此,TRIM33提供了组蛋白Kla与巨噬细胞极化之间的潜在联系。包括等温滴定量热法和蛋白质检测核磁共振在内的正交生物物理技术证实,TRIM33溴域与Kla和Kac组蛋白翻译后修饰的结合亲和力都在亚摩尔级以下。人类溴结构域的序列比对显示,在 TRIM33 结合袋中有一个独特的谷氨酸残基,与其他溴结构域相比,我们发现该残基赋予了 TRIM33 结合 Kla 的特异性。Kla 与 TRIM33 溴化结构域结合袋相互作用的分子建模和谷氨酸的定点突变证实了该残基在 TRIM33 选择性识别 Kla 中的关键作用。总之,我们的研究结果表明,TRIM33(一种含溴结构域的蛋白)是组蛋白 Kla 的新型阅读器,有可能在组蛋白 Kla 与巨噬细胞极化之间架起一座桥梁。这项研究加深了我们对组蛋白 Kla 在巨噬细胞介导的炎症中的调控作用的理解,并对其潜在的结构和生物物理机制提出了见解。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
ACS Chemical Biology
ACS Chemical Biology 生物-生化与分子生物学
CiteScore
7.50
自引率
5.00%
发文量
353
审稿时长
3.3 months
期刊介绍: ACS Chemical Biology provides an international forum for the rapid communication of research that broadly embraces the interface between chemistry and biology. The journal also serves as a forum to facilitate the communication between biologists and chemists that will translate into new research opportunities and discoveries. Results will be published in which molecular reasoning has been used to probe questions through in vitro investigations, cell biological methods, or organismic studies. We welcome mechanistic studies on proteins, nucleic acids, sugars, lipids, and nonbiological polymers. The journal serves a large scientific community, exploring cellular function from both chemical and biological perspectives. It is understood that submitted work is based upon original results and has not been published previously.
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