2J6W

R164N mutant of the RUNX1 Runt domain

Summary for 2J6W

• Entry DOI: 10.2210/pdb2j6w/pdb
• Related: 1EAN 1EAO 1EAQ 1HJB 1HJC 1IO4
• Descriptor: RUNT-RELATED TRANSCRIPTION FACTOR 1, CHLORIDE ION (3 entities in total)
• Functional Keywords: transcription factor, transcription/dna, chloride binding, acute myeloid leukemia, transcription regulation, transcription, nuclear protein, phosphorylation, aml, runx1, ig fold, runt domain, dna-binding
• Biological source: MUS MUSCULUS (MOUSE)
• Cellular location: Nucleus: Q03347
• Total number of polymer chains: 2
• Total formula weight: 31102.73

Authors

Grembecka, J.,Zhe, L.,Lukasik, S.M.,Liu, Y.,Bielnicka, I.,Bushweller, J.H.,Speck, N.A. (deposition date: 2006-10-04, release date: 2007-10-09, Last modification date: 2023-12-13)

Primary citation

Li, Z.,Lukasik, S.M.,Liu, Y.,Grembecka, J.,Bielnicka, I.,Bushweller, J.H.,Speck, N.A.
A Mutation in the S-Switch Region of the Runt Domain Alters the Dynamics of an Allosteric Network Responsible for Cbfbeta Regulation.
J.Mol.Biol., 364:1073-, 2006

PubMed Abstract: The Runt domain is the DNA binding domain of the core binding factor (CBF) Runx subunits. The CBFs are transcription factors that play critical roles in hematopoiesis, bone, and neuron development in mammals. A common non-DNA binding CBFbeta subunit heterodimerizes with the Runt domain of the Runx proteins and allosterically regulates its affinity for DNA. Previous NMR dynamics studies suggested a model whereby CBFbeta allosterically regulates DNA binding by quenching conformational exchange in the Runt domain, particularly in the S-switch region and the betaE'-F loop. We sought to test this model, and to this end introduced all possible single amino acid substitutions into the S-switch region and the betaE'-F loop, and screened for mutations that enhanced DNA-binding. We demonstrate that one Runt domain mutant, R164N, binds both DNA and CBFbeta with higher affinity, but it is less sensitive to allosteric regulation by CBFbeta. Analysis of NMR relaxation data shows that the chemical exchange exhibited by the wild-type Runt domain is largely quenched by the R164N substitution. These data support a model in which the dynamic behavior of a network of residues connecting the CBFbeta and DNA binding sites on the Runt domain plays a critical role in the mechanism of allosteric regulation. This study provides an important functional link between dynamic behavior and protein allosteric function, consistent with results on other allosterically regulated proteins.

PubMed: 17059830
DOI: 10.1016/J.JMB.2006.10.002

Experimental method

X-RAY DIFFRACTION (2.6 Å)