Matija Popovic and Hans Wienk contributed equally to this work.
The basic helix–loop–helix region of the transcriptional repressor hairy and enhancer of split 1 is preorganized to bind DNA
Article first published online: 25 JAN 2014
Copyright © 2014 Wiley Periodicals, Inc.
Proteins: Structure, Function, and Bioinformatics
Volume 82, Issue 4, pages 537–545, April 2014
How to Cite
Popovic, M., Wienk, H., Coglievina, M., Boelens, R., Pongor, S. and Pintar, A. (2014), The basic helix–loop–helix region of the transcriptional repressor hairy and enhancer of split 1 is preorganized to bind DNA. Proteins, 82: 537–545. doi: 10.1002/prot.24507
Matija Popovic's current address is Division of Molecular Structure, National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, United Kingdom.
- Issue published online: 6 MAR 2014
- Article first published online: 25 JAN 2014
- Accepted manuscript online: 8 JAN 2014 07:59PM EST
- Manuscript Accepted: 6 JAN 2014
- Manuscript Revised: 19 DEC 2013
- Manuscript Received: 30 AUG 2013
- EU (Bio-NMR, European Commission's FP7) for access to the SONNMRLSF European Large-Scale NMR Facility in Utrecht, the Netherlands. Grant Number: 261863
- secondary structure;
- backbone dynamics;
- chemical shift perturbation;
- conformational selection
Hairy and enhancer of split 1, one of the main downstream effectors in Notch signaling, is a transcriptional repressor of the basic helix–loop–helix (bHLH) family. Using nuclear magnetic resonance methods, we have determined the structure and dynamics of a recombinant protein, H1H, which includes an N-terminal segment, b1, containing functionally important phosphorylation sites, the basic region b2, required for binding to DNA, and the HLH domain. We show that a proline residue in the sequence divides the protein in two parts, a flexible and disordered N-terminal region including b1 and a structured, mainly helical region comprising b2 and the HLH domain. Binding of H1H to a double strand DNA oligonucleotide was monitored through the chemical shift perturbation of backbone amide resonances, and showed that the interaction surface involves not only the b2 segment but also several residues in the b1 and HLH regions. Proteins 2014; 82:537–545. © 2014 Wiley Periodicals, Inc.