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A CON-based NMR assignment strategy for pro-rich intrinsically disordered proteins with low signal dispersion: the C-terminal domain of histone H1.0 as a case study

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Abstract

The C-terminal domain of histone H1.0 (C-H1.0) is involved in DNA binding and is a main determinant of the chromatin condensing properties of histone H1.0. Phosphorylation at the (S/T)-P-X-(K/R) motifs affects DNA binding and is crucial for regulation of C-H1.0 function. Since C-H1.0 is an intrinsically disordered domain, solution NMR is an excellent approach to characterize the effect of phosphorylation on the structural and dynamic properties of C-H1.0. However, its very repetitive, low-amino acid-diverse and Pro-rich sequence, together with the low signal dispersion observed at the 1H–15N HSQC spectra of both non- and tri-phosphorylated C-H1.0 preclude the use of standard 1H-detected assignment strategies. We have achieved an essentially complete assignment of the heavy backbone atoms (15N, 13C′ and 13Cα), as well as 1HN and 13Cβ nuclei, of non- and tri-phosphorylated C-H1.0 by applying a novel 13C-detected CON-based strategy. No C-H1.0 region with a clear secondary structure tendency was detected by chemical shift analyses, confirming at residue level that C-H1.0 is disordered in aqueous solution. Phosphorylation only affected the chemical shifts of phosphorylated Thr’s, and their adjacent residues. Heteronuclear {1H}–15N NOEs were also essentially equal in the non- and tri-phosphorylated states. Hence, structural tendencies and dynamic properties of C-H1.0 free in aqueous solution are unmodified by phosphorylation. We propose that the assignment strategy used for C-H1.0, which is based on the acquisition of only a few 3D spectra, is an excellent choice for short-lived intrinsically disordered proteins with repetitive sequences.

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Acknowledgements

This work has been supported projects CTQ2014-52633-P and CTQ2017-84371-P from Spanish MINECO and MCIU, respectively (both co-financed by European FEDER funds). BB was a recipient of pre-doctoral FPI scholarship BES-2015-073383 from Spanish MINECO. The NMR experiments were performed in the “Manuel Rico” NMR laboratory (LMR) of the Spanish National Research Council (CSIC).

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Authors and Affiliations

  1. Departamento de Química-Física Biológica, Instituto de Química Física Rocasolano (IQFR-CSIC), Serrano 119, 28006, Madrid, Spain

    Belén Chaves-Arquero, David Pantoja-Uceda & M. Angeles Jiménez

  2. Departamento de Bioquímica y Biología Molecular, Facultad de Biociencias, Universidad Autónoma de Barcelona (UAB), Bellaterra, Barcelona, Spain

    Alicia Roque, Inmaculada Ponte & Pedro Suau

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  1. Belén Chaves-Arquero
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  2. David Pantoja-Uceda
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  4. Inmaculada Ponte
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  6. M. Angeles Jiménez
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Correspondence to M. Angeles Jiménez.

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Dedicated to Prof. Jorge Santoro on the occasion of his retirement.

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Chaves-Arquero, B., Pantoja-Uceda, D., Roque, A. et al. A CON-based NMR assignment strategy for pro-rich intrinsically disordered proteins with low signal dispersion: the C-terminal domain of histone H1.0 as a case study. J Biomol NMR 72, 139–148 (2018). https://doi.org/10.1007/s10858-018-0213-2

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