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Biomolecular NMR Assignments

, Volume 12, Issue 1, pp 183–187 | Cite as

NMR resonance assignments of RNase P protein from Thermotoga maritima

  • Danyun Zeng
  • Benjamin P. Brown
  • Markus W. Voehler
  • Sheng Cai
  • Nicholas J. Reiter
Article

Abstract

Ribonuclase P (RNase P) is an essential metallo-endonuclease that catalyzes 5′ precursor-tRNA (ptRNA) processing and exists as an RNA-based enzyme in bacteria, archaea, and eukaryotes. In bacteria, a large catalytic RNA and a small protein component assemble to recognize and accurately cleave ptRNA and tRNA-like molecular scaffolds. Substrate recognition of ptRNA by bacterial RNase P requires RNA–RNA shape complementarity, intermolecular base pairing, and a dynamic protein–ptRNA binding interface. To gain insight into the binding specificity and dynamics of the bacterial protein–ptRNA interface, we report the backbone and side chain 1H, 13C, and 15N resonance assignments of the hyperthermophilic Thermatoga maritima RNase P protein in solution at 318 K. Our data confirm the formation of a stable RNA recognition motif (RRM) with intrinsic heterogeneity at both the N- and C-terminus of the protein, consistent with available structural information. Comprehensive resonance assignments of the bacterial RNase P protein serve as an important first step in understanding how coupled RNA binding and protein–RNA conformational changes give rise to ribonucleoprotein function.

Keywords

NMR resonance assignment Thermatoga maritima RNase P Protein–tRNA binding Ribonucleoprotein TALOS-N prediction 

Notes

Acknowledgements

This work was supported in part by grants for NMR instrumentation from the NSF (0922862), NIH (S10 RR025677) and Vanderbilt University. BPB acknowledges support from the NIGMS (T32GM007347). NJR acknowledges support from Vanderbilt University and Marquette University, as well as funding from the American Heart Association (14GRNT20380334) and the NIH (GM120572). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of ChemistryMarquette UniversityMilwaukeeUSA
  2. 2.Chemical and Physical Biology ProgramVanderbilt UniversityNashvilleUSA
  3. 3.Center for Structural BiologyVanderbilt UniversityNashvilleUSA

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