The Bacterial Two-Hybrid System Uncovers the Involvement of Acetylation in Regulating of Lrp Activity in Salmonella Typhimurium.
The Bacterial Two-Hybrid System Uncovers the Involvement of Acetylation in Regulating of Lrp Activity in Salmonella Typhimurium.
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细菌双杂交系统揭示了乙酰化在鼠伤寒沙门氏菌 Lrp 活性调节中的作用。
DOI：
10.3389/fmicb.2016.01864
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发表时间：
2016
影响因子：
5.2
通讯作者：
Yao YF
中科院分区：
文献类型：
--
作者：
Qin R;Sang Y;Ren J;Zhang Q;Li S;Cui Z;Yao YF
N-𝜀-赖氨酸乙酰化是细菌中一种丰富而重要的翻译后修饰。我们使用细菌双杂交系统来筛选鼠伤寒沙门氏菌的基因组文库，以确定与乙酰基转移酶PAT-或去乙酰基酶CobB介导的乙酰化相关的潜在蛋白质。通过体外去乙酰化实验验证STM14_1074、NRDF、RHAR等潜在靶点的有效性。LRP是一种亮氨酸反应调节蛋白，也是全球调节蛋白，被证明与PAT相互作用。我们进一步证明，在体外，LRP可以被PAT乙酰化，并被NAD+依赖的Cobb去乙酰化。具体来说，LRP的螺旋-转角螺旋(HTH)DNA结合区中保守的赖氨酸残基36(K36)被乙酰化。K36的乙酰化通过改变与靶启动子的亲和力而损害LRP的功能。酵母凝集试验证实，K36基因模拟乙酰化突变提高了自身的转录水平，减弱了包括FIMA在内的LRP调控基因的mRNA水平。这些结果表明，乙酰化调节LRP的DNA结合活性，提示转录因子的乙酰化修饰是一种保守的调控方式，以调节细菌和真核生物中的基因表达。
N𝜀-lysine acetylation is an abundant and important Post-translational modification in bacteria. We used the bacterial two-hybrid system to screen the genome library of the Salmonella Typhimurium to identify potential proteins involved in acetyltransferase Pat – or deacetylase CobB-mediated acetylation. Then, the in vitro (de)acetylation assays were used to validate the potential targets, such as STM14_1074, NrdF, RhaR. Lrp, a leucine-responsive regulatory protein and global regulator, was shown to interact with Pat. We further demonstrate that Lrp could be acetylated by Pat and deacetylated by NAD+-dependent CobB in vitro. Specifically, the conserved lysine residue 36 (K36) in helix-turn-helix (HTH) DNA-binding domain of Lrp was acetylated. Acetylation of K36 impaired the function of Lrp through altering the affinity with the target promoter. The mutation of K36 in chromosome mimicking acetylation enhanced the transcriptional level of itself and attenuated the mRNA levels of Lrp-regulated genes including fimA, which was confirmed by yeast agglutination assay. These findings demonstrate that the acetylation regulates the DNA-binding activity of Lrp, suggesting that acetylation modification of transcription factors is a conserved regulatory manner to modulate gene expression in bacteria and eukaryotes.
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影响因子：
3.2
作者：
Handford, Jennifer I.;Ize, Berengere;Palmer, Tracy
通讯作者：
Palmer, Tracy
影响因子：
5.6
作者：
Babu, MM;Teichmann, SA;Aravind, L
通讯作者：
Aravind, L
DOI：
10.1073/pnas.0807227105
发表时间：
2008-12-09
影响因子：
11.1
作者：
Cho, Byung-Kwan;Barrett, Christian L.;Palsson, Bernhard O.
通讯作者：
Palsson, Bernhard O.
影响因子：
3
作者：
DUGUID, JP;CAMPBELL, I
通讯作者：
CAMPBELL, I
DOI：
10.1073/pnas.1105028108
发表时间：
2011-05-17
影响因子：
11.1
作者：
Arbely, Eyal;Natan, Eviatar;Fersht, Alan R.
通讯作者：
Fersht, Alan R.