Abstract
The PilF protein from the thermophilic bacterium Thermus thermophilus is a traffic ATPase powering the assembly of the DNA translocation machinery as well as of type 4 pili. Thereby PilF mediates the natural transformability of T. thermophilus. PilF contains a C-terminal ATPase domain and three N-terminal domains with partial homology to so-called general secretory pathway II (GSPII) domains. These three GSPII domains (GSPII-A, GSPII-B and GSPII-C) are essential for pilus assembly and twitching motility. They show varying degrees of sequence homology to the N-terminal domain of the ATPase MshE from Vibrio cholerae which binds the bacterial second messenger molecule c-di-GMP. NMR experiments demonstrate that the GSPII-B domain of PilF also binds c-di-GMP with high affinity and forms a 1:1 complex in slow exchange on the NMR time scale. As a prerequisite for structural studies of c-di-GMP binding to the GSPII-B domain of T. thermophilus PilF we present here the NMR resonance assignments for the apo and the c-di-GMP bound state of GSPII-B. In addition, we map the binding site for c-di-GMP on the GSPII-B domain using chemical shift perturbation data and compare the dynamics of the apo and the c-di-GMP-bound state of the GSPII-B domain based on {1H},15N-hetNOE data.
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Data availability
The assigned 1H, 13C, 15N chemical shifts for the apo-state and the c-di-GMP-bound-state of PilF159–302 have been deposited in the BioMagResBank (http://www.bmrb.wisc.edu/) under the Accession Numbers 27852 (apo-state) and 27853 (c-di-GMP complex).
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Acknowledgements
We are very grateful to Stefanie Düsterhus for valuable help with sample preparation and analysis. This work was supported by the Deutsche Forschungsgemeinschaft (DFG) through Grant Wo 901/7-1 to JW which is a part of the DFG special focus program SPP 1879 ‘Nucleotide Second Messenger Signaling in Bacteria’ and Grant AV9/6-2 to BA. The Center for Biomolecular Magnetic Resonance (BMRZ) at the Goethe-University Frankfurt is generously supported by the state of Hesse.
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Neißner, K., Keller, H., Duchardt-Ferner, E. et al. NMR resonance assignments for the GSPII-B domain of the traffic ATPase PilF from Thermus thermophilus in the apo and the c-di-GMP-bound state. Biomol NMR Assign 13, 383–390 (2019). https://doi.org/10.1007/s12104-019-09911-z
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DOI: https://doi.org/10.1007/s12104-019-09911-z