Traore, DAK ORCID: https://orcid.org/0000-0003-1001-4716, Wisniewski, JA, Flanigan, SF, Conroy, PJ, Panjikar, S, Mok, Y-F, Lao, C, Griffin, MDW, Adams, V, Rood, JI and Whisstock, JC (2018) Crystal structure of TcpK in complex with oriT DNA of the antibiotic resistance plasmid pCW3. Nature Communications, 9 (1).

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Abstract

Conjugation is fundamental for the acquisition of new genetic traits and the development of antibiotic resistance in pathogenic organisms. Here, we show that a hypothetical Clostridium perfringens protein, TcpK, which is encoded by the tetracycline resistance plasmid pCW3, is essential for efficient conjugative DNA transfer. Our studies reveal that TcpK is a member of the winged helix-turn-helix (wHTH) transcription factor superfamily and that it forms a dimer in solution. Furthermore, TcpK specifically binds to a nine-nucleotide sequence that is present as tandem repeats within the pCW3 origin of transfer (oriT). The X-ray crystal structure of the TcpK–TcpK box complex reveals a binding mode centered on and around the β-wing, which is different from what has been previously shown for other wHTH proteins. Structure-guided mutagenesis experiments validate the specific interaction between TcpK and the DNA molecule. Additional studies highlight that the TcpK dimer is important for specific DNA binding.

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Subjects: Q Science > Q Science (General)
Q Science > QH Natural history
Divisions: Faculty of Natural Sciences > School of Life Sciences
Depositing User: Symplectic
Date Deposited: 14 Aug 2019 08:31
Last Modified: 16 Aug 2019 13:57
URI: http://eprints.keele.ac.uk/id/eprint/6697

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