Crystallization and preliminary X-ray analysis of the Pax6 paired domain bound to the Pax6 gene enhancer.
Pax6 is a member of the Pax family of transcription factors and is essential for eye development. Pax6 has two DNA-binding domains: the paired domain and the homeodomain. The Pax6 paired domain is involved in Pax6 gene autoregulation by binding to its enhancer. In this study, crystallization and preliminary X-ray diffraction analysis of the mammalian Pax6 paired domain in complex with the Pax6 gene enhancer was attempted. The Pax6 paired domain complexed with an optimized 25 bp DNA fragment was crystallized by the hanging-drop vapour-diffusion method. The crystal diffracted synchrotron radiation to 3.0/3.7 A resolution and belongs to the monoclinic space group P2(1), with unit-cell parameters a = 62.21, b = 70.69, c = 176.03 A, beta = 90.54 degrees. Diffraction data were collected to 3.7 A resolution.
Ito
Makoto
M
Department of Structural Biology, Biomolecular Engineering Research Institute, 6-2-3 Furuedai, Suita, Osaka 565-0874, Japan. morikawa@beri.or.jp
Oyama
Takuji
T
Okazaki
Kenji
K
Morikawa
Kosuke
K
D001665:Binding Sites; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004247:DNA / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D030562:Databases, Protein; D004058:Diffusion; D004742:Enhancer Elements, Genetic; D004926:Escherichia coli / Q000378:metabolism; D005136:Eye Proteins / Q000737:chemistry* / Q000235:genetics*; D005786:Gene Expression Regulation; D018398:Homeodomain Proteins / Q000737:chemistry* / Q000235:genetics*; D006801:Humans; D008968:Molecular Conformation; D009841:Oligonucleotides / Q000737:chemistry; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000737:chemistry* / Q000235:genetics*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012097:Repressor Proteins / Q000737:chemistry* / Q000235:genetics*; D017356:Synchrotrons; D014961:X-Ray Diffraction
D018076:DNA, Complementary; D005136:Eye Proteins; D018398:Homeodomain Proteins; D009841:Oligonucleotides; D000071841:PAX6 Transcription Factor; C000605730:PAX6 protein, human; D051761:Paired Box Transcription Factors; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D004247:DNA