Structural organization of Staf-DNA complexes.
The transactivator Staf, which contains seven contiguous zinc fingers of the C(2)-H(2)type, exerts its effects on gene expression by binding to specific targets in vertebrate small nuclear RNA (snRNA) and snRNA-type gene promoters. Here, we have investigated the interaction of the Staf zinc finger domain with the optimal Xenopus selenocysteine tRNA (xtRNA(Sec)) and human U6 snRNA (hU6) Staf motifs. Generation of a series of polypeptides containing increasing numbers of Staf zinc fingers tested in binding assays, by interference techniques and by binding site selection served to elucidate the mode of interaction between the zinc fingers and the Staf motifs. Our results provide strong evidence that zinc fingers 3-6 represent the minimal zinc finger region for high affinity binding to Staf motifs. Furthermore, we show that the binding of Staf is achieved through a broad spectrum of close contacts between zinc fingers 1-6 and xtRNA(Sec)or optimal sites or between zinc fingers 3-6 and the hU6 site. Extensive DNA major groove contacts contribute to the interaction with Staf that associates more closely with the non-template than with the template strand. Based on these findings and the structural information provided by the solved structures of other zinc finger-DNA complexes, we propose a model for the interaction between Staf zinc fingers and the xtRNA(Sec), optimal and hU6 sites.
Schaub
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M
UPR 9002 du CNRS 'Structure des Macromolécules Biologiques et Mécanismes de Reconnaissance', IBMC, 15 rue René Descartes, 67084 Strasbourg Cedex, France.
Krol
A
A
Carbon
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P
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D012342:RNA, Small Nuclear / Q000235:genetics*; D012345:RNA, Transfer, Amino Acid-Specific / Q000235:genetics*; D013698:Templates, Genetic; D015534:Trans-Activators / Q000737:chemistry* / Q000378:metabolism*; D014714:Vertebrates; D029867:Xenopus Proteins; D014982:Xenopus laevis; D016335:Zinc Fingers
D004268:DNA-Binding Proteins; D012342:RNA, Small Nuclear; D012345:RNA, Transfer, Amino Acid-Specific; D015534:Trans-Activators; D029867:Xenopus Proteins; C114253:ZNF143 protein, human; C036928:tRNA, selenocysteine-; C491943:znf143 protein, Xenopus