title: Solution structure of the Oct-1 POU homeodomain determined by NMR and restrained molecular dynamics.
abstract: The POU homeodomain (POUhd), a divergent member of the well-studied class of homeodomain proteins, is the C-terminal part of the bipartite POU domain, the conserved DNA-binding domain of the POU proteins. In this paper we present the solution structure of POUhd of the human Oct-1 transcription factor. This fragment was overexpressed in Escherichia coli and studied by two- and three-dimensional homo- and heteronuclear NMR techniques, resulting in virtually complete 1H and 15N resonance assignments for residues 2-60. Using distance and dihedral constraints derived from the NMR data, 50 distance geometry structures were calculated, which were refined by means of restrained molecular dynamics. From this set a total of 31 refined structures were selected, having low constraint energy and few constraint violations. The ensemble of 31 structures displays a root-mean-square deviation of the coordinates of 0.59 A with respect to the average structure, calculated over the backbone atoms of residues 6 to 54. The fold of POUhd is very similar to that of the canonical homeodomains. Interestingly, the recognition helix of the free POUhd ends at residue 53, while in the cocrystal structure of the intact POU domain with the DNA octamer motif [Klemm, J.D., Rould, M.A., Aurora, R., Herr, W. and Pabo, C.O. (1994) Cell, 77, 21-32] this helix in the POUhd subdomain is extended as far as residue 60.
authors.affiliation: Bijvoet Center for Biomolecular Research, Utrecht University, The Netherlands.
authors.forename: M
authors.initials: M
authors.lastname: Cox
authors.forename: P J
authors.initials: PJ
authors.lastname: van Tilborg
authors.forename: W
authors.initials: W
authors.lastname: de Laat
authors.forename: R
authors.initials: R
authors.lastname: Boelens
authors.forename: H C
authors.initials: HC
authors.lastname: van Leeuwen
authors.forename: P C
authors.initials: PC
authors.lastname: van der Vliet
authors.forename: R
authors.initials: R
authors.lastname: Kaptein
mesh_terms: D000595:Amino Acid Sequence; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018398:Homeodomain Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051863:Host Cell Factor C1; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D050811:Octamer Transcription Factor-1; D050809:POU Domain Factors; D017433:Protein Structure, Secondary; D012996:Solutions; D013816:Thermodynamics; D014157:Transcription Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; C497692:HCFC1 protein, human; D018398:Homeodomain Proteins; D051863:Host Cell Factor C1; D050811:Octamer Transcription Factor-1; D050809:POU Domain Factors; C494165:POU2F1 protein, human; D012996:Solutions; D014157:Transcription Factors; D004247:DNA