Nε-lysine acetylation of a bacterial transcription factor inhibits Its DNA-binding activity.

Nε-lysine acetylation of a bacterial transcription factor inhibits Its DNA-binding activity.
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DOI:
10.1371/journal.pone.0015123
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发表时间:
2010-12-31
期刊:
影响因子:
3.7
通讯作者:
Escalante-Semerena JC
Escalante-Semerena JC
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Thao S;Chen CS;Zhu H;Escalante-Semerena JC

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据报道,有证据表明真核生物和古细菌使用可逆的 N ε-赖氨酸 (N ε-Lys) 乙酰化来调节基因表达,但缺乏细菌利用 N ε-Lys 乙酰化用于此目的的证据。在这里,我们报告的数据支持细菌可以通过调节转录因子(TF)的乙酰化状态来控制基因表达的观点。我们筛选了大肠杆菌蛋白质组中细菌 Gcn5 样蛋白乙酰转移酶 (Pat) 的底物。 Pat 乙酰化了四种转录因子,包括 RcsB 全局调节蛋白,它控制许多细菌的细胞分裂、荚膜和鞭毛生物合成。 Pat 乙酰化 RcsB 的残基 Lys180,以及 NAD+ 依赖性 Sir2 (sirtuin) 样蛋白脱乙酰酶 (CobB) 脱乙酰乙酰化 RcsB (RcsBAc),证明 RcsB 的 N ε-Lys 乙酰化是可逆的。对在 Lys180 处进行取代的 RcsBAc 和变体 RcsB 蛋白的分析提供了生化和生理学证据,表明 Lys180 是 RcsB DNA 结合活性的关键残基。这些发现进一步证明转录因子的可逆 N ε-Lys 乙酰化是所有细胞使用的基因表达调节模式。
Evidence suggesting that eukaryotes and archaea use reversible N ε-lysine (N ε-Lys) acetylation to modulate gene expression has been reported, but evidence for bacterial use of N ε-Lys acetylation for this purpose is lacking. Here, we report data in support of the notion that bacteria can control gene expression by modulating the acetylation state of transcription factors (TFs). We screened the E. coli proteome for substrates of the bacterial Gcn5-like protein acetyltransferase (Pat). Pat acetylated four TFs, including the RcsB global regulatory protein, which controls cell division, and capsule and flagellum biosynthesis in many bacteria. Pat acetylated residue Lys180 of RcsB, and the NAD+-dependent Sir2 (sirtuin)-like protein deacetylase (CobB) deacetylated acetylated RcsB (RcsBAc), demonstrating that N ε-Lys acetylation of RcsB is reversible. Analysis of RcsBAc and variant RcsB proteins carrying substitutions at Lys180 provided biochemical and physiological evidence implicating Lys180 as a critical residue for RcsB DNA-binding activity. These findings further the likelihood that reversible N ε-Lys acetylation of transcription factors is a mode of regulation of gene expression used by all cells.
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