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Type: Journal article
Title: Native mass spectrometry identifies an alternative DNA-binding pathway for BirA from Staphylococcus aureus
Author: Satiaputra, J.
Sternicki, L.M.
Hayes, A.J.
Pukala, T.L.
Booker, G.W.
Shearwin, K.E.
Polyak, S.W.
Citation: Scientific Reports, 2019; 9(1):2767-1-2767-13
Publisher: Springer Nature
Issue Date: 2019
ISSN: 2045-2322
Statement of
Jiulia Satiaputra, Louise M. Sternicki, Andrew J. Hayes, Tara L. Pukala, Grant W. Booker, Keith E. Shearwin, Steven W. Polyak
Abstract: An adequate supply of biotin is vital for the survival and pathogenesis of Staphylococcus aureus. The key protein responsible for maintaining biotin homeostasis in bacteria is the biotin retention protein A (BirA, also known as biotin protein ligase). BirA is a bi-functional protein that serves both as a ligase to catalyse the biotinylation of important metabolic enzymes, as well as a transcriptional repressor that regulates biotin biosynthesis, biotin transport and fatty acid elongation. The mechanism of BirA regulated transcription has been extensively characterized in Escherichia coli, but less so in other bacteria. Biotin-induced homodimerization of E. coli BirA (EcBirA) is a necessary prerequisite for stable DNA binding and transcriptional repression. Here, we employ a combination of native mass spectrometry, in vivo gene expression assays, site-directed mutagenesis and electrophoretic mobility shift assays to elucidate the DNA binding pathway for S. aureus BirA (SaBirA). We identify a mechanism that differs from that of EcBirA, wherein SaBirA is competent to bind DNA as a monomer both in the presence and absence of biotin and/or MgATP, allowing homodimerization on the DNA. Bioinformatic analysis demonstrated the SaBirA sequence used here is highly conserved amongst other S. aureus strains, implying this DNA-binding mechanism is widely employed.
Keywords: Staphylococcus aureus; Biotin; Carbon-Nitrogen Ligases; Bacterial Proteins; Escherichia coli Proteins; Repressor Proteins; DNA; Spectrometry, Mass, Electrospray Ionization; Amino Acid Sequence; Protein Structure, Tertiary; Protein Binding; Dimerization; Nanotechnology
Rights: © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit
RMID: 0030110411
DOI: 10.1038/s41598-019-39398-6
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Appears in Collections:Molecular and Biomedical Science publications

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