N−Lysine Acetylation of a Bacterial Transcription Factor Inhibits Its DNA-Binding Activity

Baylor College of Medicine, United States of America
PLoS ONE (Impact Factor: 3.23). 12/2010; 5(12):e15123. DOI: 10.1371/journal.pone.0015123
Source: PubMed


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 (RcsB(Ac)), demonstrating that N(ε)-Lys acetylation of RcsB is reversible. Analysis of RcsB(Ac) 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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Available from: Jorge C Escalante-Semerena,
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    • "The LysAc of the response regulator RcsB, which regulates the transcription of more than 150 genes in E. coli, is a well-known example. Thus, in vitro acetylation of a Lys residue that belongs to the DNA-binding, helix-turn-helix motif of RcsB seems to alter its DNA-binding activity [74]. Recently, Hu et al [75] showed that in vivo acetylation of this protein reduces the transcription of the small non-encoding RNA (rprA), which activates the translation of stationary-phase sigma factor RpoS [76]. "
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    ABSTRACT: Post-translational modifications of proteins are key events in cellular metabolism and physiology regulation. Lysine acetylation is one of the best studied protein modifications in eukaryotes, but, until recently, ignored in bacteria. However, proteomic advances have highlighted the diversity of bacterial lysine-acetylated proteins. The current data support the implication of lysine acetylation in various metabolic pathways, adaptation and virulence. In this review, we present a broad overview of the current knowledge of lysine acetylation in bacteria. We emphasize particularly the significant contribution of proteomics in this field. This article is protected by copyright. All rights reserved.
    Proteomics 09/2015; DOI:10.1002/pmic.201500258 · 3.81 Impact Factor
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    • "In general, MS-based strategies allow the non-biased identification of a high number of targeted proteins/pathways. Other techniques , such as protein microarrays, have also been used for the identification of substrates of acetyltransferases, although in vitro specificities may not mirror what happens in vivo [47]. "
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    ABSTRACT: Post-translational modification of proteins is a reversible mechanism of cellular adaptation to changing environmental conditions. In eukaryotes, the physiological relevance of N-ɛ-lysine protein acetylation is well demonstrated. In recent times, important roles in the regulation of metabolic processes in bacteria are being uncovered, adding complexity to cellular regulatory networks. The aim of this mini-review is to sum up the current state-of-the-art in the regulation of bacterial physiology by protein acetylation. Current knowledge on the molecular biology aspects of known bacterial protein acetyltransferases and deacetylases will be summarized. Protein acetylation in Escherichia coli, Salmonella enterica, Bacillus subtilis, Rhodopseudomonas palustris and Mycobacterium tuberculosis, will be explained in the light of their physiological relevance. Progress in the elucidation of bacterial acetylomes and the emerging understanding of chemical acylation mechanisms will be discussed together with their regulatory and evolutionary implications. Fundamental molecular studies detailing this recently discovered regulatory mechanism pave the way for their prospective application for the construction of synthetic regulation networks.
    New Biotechnology 12/2014; 31(6). DOI:10.1016/j.nbt.2014.03.002 · 2.90 Impact Factor
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    • "In a second study, the influence of Nε-L-lysine acetylation on the activity of transcription factors in E. coli was investigated (Thao et al., 2010). A proteome screen for substrates of the Gcn5-like protein acetyltransferase (Pat) afforded four transcription factors as substrates. "
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    ABSTRACT: The expansion of the genetic code with non-canonical amino acids (ncAA) enables the chemical and biophysical properties of proteins to be tailored, inside cells, with a previously unattainable level of precision. A wide range of ncAA with functions not found in canonical amino acids have been genetically encoded in recent years and have delivered insights into biological processes that would be difficult to access with traditional approaches of molecular biology. A major field for the development and application of novel ncAA-functions has been transcription and its regulation. This is particularly attractive, since advanced DNA sequencing- and proteomics-techniques continue to deliver vast information on these processes on a global level, but complementing methodologies to study them on a detailed, molecular level and in living cells have been comparably scarce. In a growing number of studies, genetic code expansion has now been applied to precisely control the chemical properties of transcription factors, RNA polymerases and histones, and this has enabled new insights into their interactions, conformational changes, cellular localizations and the functional roles of posttranslational modifications.
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