Native mass spectrometry identifies an alternative DNA-binding pathway for BirA from Staphylococcus aureus
| العنوان: | Native mass spectrometry identifies an alternative DNA-binding pathway for BirA from Staphylococcus aureus |
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| المؤلفون: | Tara L. Pukala, Andrew J. Hayes, Louise M. Sternicki, Keith E. Shearwin, Jiulia Satiaputra, Steven W. Polyak, Grant W. Booker |
| المساهمون: | Satiaputra, Jiulia, Sternicki, Louise M, Hayes, Andrew J, Pukala, Tara L, Booker, Grant W, Shearwin, Keith E, Polyak, Steven W |
| المصدر: | Scientific Reports, Vol 9, Iss 1, Pp 1-13 (2019) Scientific Reports |
| بيانات النشر: | Nature Publishing Group, 2019. |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | 0301 basic medicine, Spectrometry, Mass, Electrospray Ionization, Staphylococcus aureus, Repressor, Biotin, lcsh:Medicine, Article, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Bacterial Proteins, biotin, Nanotechnology, Carbon-Nitrogen Ligases, Amino Acid Sequence, lcsh:Science, chemistry.chemical_classification, DNA ligase, Multidisciplinary, biology, Escherichia coli Proteins, lcsh:R, DNA, Protein Structure, Tertiary, Repressor Proteins, Biotin transport, 030104 developmental biology, Biochemistry, chemistry, Biotinylation, biology.protein, Fatty acid elongation, lcsh:Q, Protein A, Dimerization, 030217 neurology & neurosurgery, Protein Binding |
| الوصف: | 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. |
| اللغة: | English |
| تدمد: | 2045-2322 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::e872c44a140fdec1fb6afa9987e76fd7 http://link.springer.com/article/10.1038/s41598-019-39398-6 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....e872c44a140fdec1fb6afa9987e76fd7 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 20452322 |
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