Crystallization of the Oct-1/SNAP190 peptide/DNA complex.
Crystals of the Oct-1 POU/SNAP190 peptide/DNA tertiary complex have been obtained by hanging-drop vapor diffusion at 293K in 20% 2-propanol, 20% PEG 4000 and 0.1M sodium citrate pH 5.6. The Oct-1 POU protein has two domains, one a homeodomain and the other a POU domain, which are connected by a flexible linker. The DNA used in the complex is slightly different in the octamer region compared with the two previously crystallized Oct-1 POU/DNA complexes. The DNA is 14 base pairs, with an octamer sequence of 5'-ATGTAGAT-3' and an overhang of one base on both strands. The SNAP190 peptide is 27 amino acids long (residues 884-910). The crystals diffract to 2.3 A (94.1% completeness) at the synchrotron under cryogenic (123K) conditions. The crystals are triclinic, space group P1, with unit-cell parameters a = 36.4, b = 54.9, c = 77.6A, alpha = 94.9, beta = 99.6, gamma = 109.2 degrees. This structure will provide insight into how Oct-1 interacts with SNAP190, a critical component of the small nuclear RNA-activating protein complex (SNAPc). Transcription of human snRNA genes is activated by these direct protein-protein interactions.
Hovde
Stacy
S
Michigan State University Chemistry Department, East Lansing, MI 48824, USA.
Brooks
Aimee
A
Strong
Katie
K
Geiger
James H
JH
D001426:Bacterial Proteins / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D004247:DNA / Q000737:chemistry*; D004268:DNA-Binding Proteins / Q000737:chemistry*; D004926:Escherichia coli / Q000737:chemistry*; D051863:Host Cell Factor C1; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D050811:Octamer Transcription Factor-1; D010446:Peptide Fragments / Q000737:chemistry; D011487:Protein Conformation; D014157:Transcription Factors / Q000737:chemistry*
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D051863:Host Cell Factor C1; D050811:Octamer Transcription Factor-1; D010446:Peptide Fragments; C116351:SNAPC4 protein, human; D014157:Transcription Factors; D004247:DNA