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Resonance assignments and secondary structure of thermophile single‐stranded DNA binding protein from Sulfolobus solfataricus at 323K

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Abstract

Single-stranded DNA (ssDNA)-binding proteins (SSBs) are essential for DNA replication, recombination, and repair processes in all organisms. Sulfolobus solfataricus (S. solfataricus), a hyperthermophilic species, overexpresses its SSB (S. solfataricus SSB (SsoSSB)) to protect ssDNA during DNA metabolisms. Even though the crystal structure of apo SsoSSB and its ssDNA-bound solution structure have been reported at room temperature, structural information at high temperature is not yet available. To find out how SsoSSB maintains its structure and ssDNA binding affinity at high temperatures, we performed multidimensional NMR experiments for SsoSSB at 323K. In this study, we present the backbone and side chain chemical shifts and predict the secondary structure of SsoSSB from the chemical shifts. We found that SsoSSB is ordered, even at high temperatures, and has the same fold at high temperature as at room temperature. Our data will help improve structural analyses and our understanding of the features of thermophilic proteins.

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Acknowledgements

We thank GIST Central Research Facilities (GIST, Gwangju) for allowing us to use their NMR spectrometer. This work was supported by the National Research Foundation (NRF) of Korea [Grant No. 2018R1A2B6004388 to C.-J.P], which is funded by the Korean government (MSIT); by a GIST Research Institute grant funded by GIST in 2020; the National Science Foundation (NSF) [DBI-2051595 to W.L.] and the University of Colorado Denver [the start-up support to W.L.].

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Authors and Affiliations

  1. Department of Chemistry, Gwangju Institute of Science and Technology, Gwangju, 61005, Korea

    Min June Yang & Chin-Ju Park

  2. Department of Chemistry, University of Colorado Denver, Denver, CO, 80217-3364, USA

    Woonghee Lee

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  1. Min June Yang
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  2. Woonghee Lee
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  3. Chin-Ju Park
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Contributions

CP and WL designed the research. MY performed the experiments. MY and WL analyzed the data. MY, WL, and CP wrote the paper.

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Correspondence to Woonghee Lee or Chin-Ju Park.

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Yang, M.J., Lee, W. & Park, CJ. Resonance assignments and secondary structure of thermophile single‐stranded DNA binding protein from Sulfolobus solfataricus at 323K. Biomol NMR Assign 15, 159–164 (2021). https://doi.org/10.1007/s12104-020-09999-8

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