title: The Bacterial Two-Hybrid System Uncovers the Involvement of Acetylation in Regulating of Lrp Activity inSalmonellaTyphimurium.
abstract: 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 theSalmonellaTyphimurium to identify potential proteins involved in acetyltransferase Pat - or deacetylase CobB-mediated acetylation. Then, thein 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 CobBin 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 includingfimA, 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.
authors.affiliation: Key Laboratory of Agricultural Environmental Microbiology, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University Nanjing, China.
authors.forename: Ran
authors.initials: R
authors.lastname: Qin
authors.affiliation: Laboratory of Bacterial Pathogenesis, Department of Microbiology and Immunology, Institutes of Medical Sciences, Shanghai Jiao Tong University School of Medicine Shanghai, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Sang
authors.affiliation: Laboratory of Bacterial Pathogenesis, Department of Microbiology and Immunology, Institutes of Medical Sciences, Shanghai Jiao Tong University School of Medicine Shanghai, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Ren
authors.affiliation: Laboratory of Bacterial Pathogenesis, Department of Microbiology and Immunology, Institutes of Medical Sciences, Shanghai Jiao Tong University School of Medicine Shanghai, China.
authors.forename: Qiufen
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Laboratory of Bacterial Pathogenesis, Department of Microbiology and Immunology, Institutes of Medical Sciences, Shanghai Jiao Tong University School of Medicine Shanghai, China.
authors.forename: Shuxian
authors.initials: S
authors.lastname: Li
authors.affiliation: Key Laboratory of Agricultural Environmental Microbiology, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University Nanjing, China.
authors.forename: Zhongli
authors.initials: Z
authors.lastname: Cui
authors.affiliation: Laboratory of Bacterial Pathogenesis, Department of Microbiology and Immunology, Institutes of Medical Sciences, Shanghai Jiao Tong University School of MedicineShanghai, China; Department of Laboratory Medicine, Shanghai East Hospital, Tongji University School of MedicineShanghai, China.
authors.forename: Yu-Feng
authors.initials: YF
authors.lastname: Yao
keywords: DNA-binding; Lrp; Salmonella Typhimurium; bacterial two-hybrid; lysine acetylation