Swapping of the beta-hairpin region between Sp1 and GLI zinc fingers: significant role of the beta-hairpin region in DNA binding properties of C2H2-type zinc finger peptides.
The recent design strategy of zinc finger peptides has mainly focused on the alpha-helix region, which plays a direct role in DNA recognition. On the other hand, the study of non-DNA-contacting regions is extremely scarce. By swapping the beta-hairpin regions between the Sp1 and GLI zinc fingers, in this study, we investigated how the beta-hairpin region of the C(2)H(2)-type zinc finger peptides contributes to the DNA binding properties. Surprisingly, the Sp1 mutant with the GLI-type beta-hairpin had a higher DNA binding affinity than that of the wild-type Sp1. The result of the DNase I footprinting analyses also showed the change in the DNA binding pattern. In contrast, the GLI zinc finger completely lost DNA binding ability as a result of exchanging the beta-hairpin region. These results strongly indicate that the beta-hairpin region appears to function as a scaffold and has an important effect on the DNA binding properties of the C(2)H(2)-type zinc finger peptides.
Shiraishi
Yasuhisa
Y
Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.
Imanishi
Miki
M
Morisaki
Tatsuya
T
Sugiura
Yukio
Y
D000595:Amino Acid Sequence; D002942:Circular Dichroism; D018983:DNA Footprinting; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics; D003850:Deoxyribonuclease I; D006801:Humans; D006860:Hydrogen Bonding; D008969:Molecular Sequence Data; D015513:Oncogene Proteins / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics; D016133:Polymerase Chain Reaction; D011485:Protein Binding / Q000235:genetics; D011487:Protein Conformation; D017433:Protein Structure, Secondary / Q000235:genetics; D016329:Sp1 Transcription Factor / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics; D015534:Trans-Activators; D014157:Transcription Factors / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics; D000071676:Zinc Finger Protein GLI1; D016335:Zinc Fingers / Q000235:genetics
D004268:DNA-Binding Proteins; D015513:Oncogene Proteins; D010446:Peptide Fragments; D016329:Sp1 Transcription Factor; D015534:Trans-Activators; D014157:Transcription Factors; D000071676:Zinc Finger Protein GLI1; D003850:Deoxyribonuclease I