2VI6: Crystal Structure Of The Nanog Homeodomain

Citation:
Abstract
The transcription factor Nanog is an upstream regulator in early mammalian development and a key determinant of pluripotency in embryonic stem cells. Nanog binds to promoter elements of hundreds of target genes and regulates their expression by an as yet unknown mechanism. Here, we report the crystal structure of the murine Nanog homeodomain (HD) and analysis of its interaction with a DNA element derived from the Tcf3 promoter. Two Nanog amino acid pairs, unique among HD sequences, appear to affect the mechanism of nonspecific DNA recognition as well as maintain the integrity of the structural scaffold. To assess selective DNA recognition by Nanog, we performed electrophoretic mobility shift assays using a panel of modified DNA binding sites and found that Nanog HD preferentially binds the TAAT(G/T)(G/T) motif. A series of rational mutagenesis experiments probing the role of six variant residues of Nanog on its DNA binding function establish their role in affecting binding affinity but not binding specificity. Together, the structural and functional evidence establish Nanog as a distant member of a Q50-type HD despite having considerable variation at the sequence level.
PDB ID: 2VI6Download
MMDB ID: 61857
PDB Deposition Date: 2007/11/28
Updated in MMDB: 2012/09
Experimental Method:
x-ray diffraction
Resolution: 2.6  Å
Source Organism:
Similar Structures:
Biological Unit for 2VI6: monomeric; determined by author and by software (PQS)
Molecular Components in 2VI6
Label Count Molecule
Protein (1 molecule)
1
Homeobox Protein Nanog(Gene symbol: Nanog)
Molecule annotation
* Click molecule labels to explore molecular sequence information.

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