Flexible zinc finger requirement for binding of the transcriptional activator staf to U6 small nuclear RNA and tRNA(Sec) promoters.
The transactivator Staf, which contains seven zinc finger motifs, exerts its effect on gene expression by binding to specific targets in small nuclear RNA (snRNA) and snRNA-type gene promoters. In this work, binding site selection allowed us to identify the 21-base pair ATTACCCATAATGCATYGCGG sequence as the high affinity consensus binding site for Staf. It shows a high sequence divergence with Staf-responsive elements in the Xenopus selenocysteine tRNA (tRNA(Sec)) and human U6 snRNA promoters. By using a combination of approaches, we analyzed the interaction of wild-type and truncated Staf zinc finger domains with the consensus, Xenopus tRNA(Sec), and human U6 sites. Two main conclusions emerged from our data. First, the data clearly indicate that zinc finger 7 does not establish base-specific contacts in Staf-DNA complexes. The second conclusion concerns zinc finger 1, which is required for the binding to the Xenopus tRNA(Sec) site but is dispensable in the case of the human U6 site. Taking into account the sequence differences in the two sites, these findings demonstrate that Staf utilizes zinc finger 1 in a rather flexible manner, illustrating how a protein can interact with DNAs containing targets of different sequences.
Schaub
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UPR 9002 du CNRS Structure des Macromolécules Biologiques et Mécanismes de Reconnaissance, IBMC, 15 rue René Descartes, 67084 Strasbourg Cedex, France.
Krol
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A
Carbon
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P
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000494:pharmacology; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012342:RNA, Small Nuclear / Q000235:genetics*; D012345:RNA, Transfer, Amino Acid-Specific / Q000235:genetics*; D015534:Trans-Activators / Q000737:chemistry / Q000378:metabolism*; D014981:Xenopus; D029867:Xenopus Proteins; 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; D003850:Deoxyribonuclease I