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2000
Schaub M, Krol A, Carbon P
Structural organization of Staf-DNA complexes Article de journal
Dans: Nucleic Acids Res, vol. 28, no. 10, p. 2114-2121, 2000, ISBN: 10773080, (1362-4962 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acid Sequence Animals Base Sequence Binding Sites DNA-Binding Proteins/*chemistry/*metabolism Human Models, Amino Acid-Specific/*genetics Support, Genetic Trans-Activators/*chemistry/*metabolism Vertebrates Xenopus laevis Zinc Fingers, Molecular Molecular Sequence Data Nucleic Acid Conformation Plasmids/*chemistry/*metabolism Protein Conformation RNA, Non-U.S. Gov't Templates, Small Nuclear/*genetics RNA, Transfer, Unité ARN
@article{,
title = {Structural organization of Staf-DNA complexes},
author = {M Schaub and A Krol and P Carbon},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10773080},
isbn = {10773080},
year = {2000},
date = {2000-01-01},
journal = {Nucleic Acids Res},
volume = {28},
number = {10},
pages = {2114-2121},
abstract = {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.},
note = {1362-4962
Journal Article},
keywords = {Amino Acid Sequence Animals Base Sequence Binding Sites DNA-Binding Proteins/*chemistry/*metabolism Human Models, Amino Acid-Specific/*genetics Support, Genetic Trans-Activators/*chemistry/*metabolism Vertebrates Xenopus laevis Zinc Fingers, Molecular Molecular Sequence Data Nucleic Acid Conformation Plasmids/*chemistry/*metabolism Protein Conformation RNA, Non-U.S. Gov't Templates, Small Nuclear/*genetics RNA, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
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.
1999
Schaub M, Myslinski E, Krol A, Carbon P
Maximization of selenocysteine tRNA and U6 small nuclear RNA transcriptional activation achieved by flexible utilization of a Staf zinc finger Article de journal
Dans: J Biol Chem, vol. 274, no. 35, p. 25042-25050, 1999, ISBN: 10455183, (0021-9258 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acid Sequence Animals Binding Sites/genetics DNA/genetics DNA-Binding Proteins/*genetics Human Hydroxyl Radical/metabolism Microinjections Molecular Sequence Data Mutation Oocytes Peptide Fragments/immunology Promoter Regions (Genetics) Protein Binding RNA, Amino Acid-Specific/*genetics Sequence Homology Support, Genetic Trans-Activation (Genetics)/*genetics Trans-Activators/*genetics Transcription Factors/genetics Xenopus Zinc Fingers/*genetics, Non-U.S. Gov't Templates, Small Nuclear/*genetics RNA, Transfer, Unité ARN
@article{,
title = {Maximization of selenocysteine tRNA and U6 small nuclear RNA transcriptional activation achieved by flexible utilization of a Staf zinc finger},
author = {M Schaub and E Myslinski and A Krol and P Carbon},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10455183},
isbn = {10455183},
year = {1999},
date = {1999-01-01},
journal = {J Biol Chem},
volume = {274},
number = {35},
pages = {25042-25050},
abstract = {Transcriptional activators Staf and Oct-1 play critical roles in the activation of small nuclear RNA (snRNA) and snRNA-type gene transcription. Recently, we established that Staf binding to the human U6 snRNA (hU6) and Xenopus selenocysteine tRNA (xtRNA(Sec)) genes requires different sets of the seven C2-H2 zinc fingers. In this work, using a combination of oocyte microinjection, electrophoretic mobility shift assays, and missing nucleoside experiments with wild-type and mutant promoters, we demonstrate that the hU6 gene requires zinc fingers 2-7 for Staf binding and Oct-1 for maximal transcriptional activity. In contrast, the xtRNA(Sec) gene needs the binding of the seven Staf zinc fingers, but not Oct-1, for optimal transcriptional capacity. Mutation in the binding site for Staf zinc finger 1 in the tRNA(Sec) promoter reduced both Staf binding and transcriptional activity. Conversely, introduction of a zinc finger 1 binding site in the hU6 promoter increased Staf binding but interfered with the simultaneous Staf and Oct-1 binding, thus reducing transcriptional activity. Collectively, these results show that the differential utilization of Staf zinc finger 1 represents a new, critical determinant of the transcriptional activation mechanism for the Xenopus tRNA(Sec) and human U6 snRNA genes.},
note = {0021-9258
Journal Article},
keywords = {Amino Acid Sequence Animals Binding Sites/genetics DNA/genetics DNA-Binding Proteins/*genetics Human Hydroxyl Radical/metabolism Microinjections Molecular Sequence Data Mutation Oocytes Peptide Fragments/immunology Promoter Regions (Genetics) Protein Binding RNA, Amino Acid-Specific/*genetics Sequence Homology Support, Genetic Trans-Activation (Genetics)/*genetics Trans-Activators/*genetics Transcription Factors/genetics Xenopus Zinc Fingers/*genetics, Non-U.S. Gov't Templates, Small Nuclear/*genetics RNA, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Transcriptional activators Staf and Oct-1 play critical roles in the activation of small nuclear RNA (snRNA) and snRNA-type gene transcription. Recently, we established that Staf binding to the human U6 snRNA (hU6) and Xenopus selenocysteine tRNA (xtRNA(Sec)) genes requires different sets of the seven C2-H2 zinc fingers. In this work, using a combination of oocyte microinjection, electrophoretic mobility shift assays, and missing nucleoside experiments with wild-type and mutant promoters, we demonstrate that the hU6 gene requires zinc fingers 2-7 for Staf binding and Oct-1 for maximal transcriptional activity. In contrast, the xtRNA(Sec) gene needs the binding of the seven Staf zinc fingers, but not Oct-1, for optimal transcriptional capacity. Mutation in the binding site for Staf zinc finger 1 in the tRNA(Sec) promoter reduced both Staf binding and transcriptional activity. Conversely, introduction of a zinc finger 1 binding site in the hU6 promoter increased Staf binding but interfered with the simultaneous Staf and Oct-1 binding, thus reducing transcriptional activity. Collectively, these results show that the differential utilization of Staf zinc finger 1 represents a new, critical determinant of the transcriptional activation mechanism for the Xenopus tRNA(Sec) and human U6 snRNA genes.
Schaub M, Krol A, Carbon P
Flexible zinc finger requirement for binding of the transcriptional activator staf to U6 small nuclear RNA and tRNA(Sec) promoters Article de journal
Dans: J Biol Chem, vol. 274, no. 34, p. 24241-24249, 1999, ISBN: 10446199, (0021-9258 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acid Sequence Animals Binding Sites DNA-Binding Proteins/chemistry/*metabolism Deoxyribonuclease I/pharmacology Human Molecular Sequence Data *Promoter Regions (Genetics) RNA, Amino Acid-Specific/*genetics Support, Non-U.S. Gov't Trans-Activators/chemistry/*metabolism Xenopus *Zinc Fingers, Small Nuclear/*genetics RNA, Transfer, Unité ARN
@article{,
title = {Flexible zinc finger requirement for binding of the transcriptional activator staf to U6 small nuclear RNA and tRNA(Sec) promoters},
author = {M Schaub and A Krol and P Carbon},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10446199},
isbn = {10446199},
year = {1999},
date = {1999-01-01},
journal = {J Biol Chem},
volume = {274},
number = {34},
pages = {24241-24249},
abstract = {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.},
note = {0021-9258
Journal Article},
keywords = {Amino Acid Sequence Animals Binding Sites DNA-Binding Proteins/chemistry/*metabolism Deoxyribonuclease I/pharmacology Human Molecular Sequence Data *Promoter Regions (Genetics) RNA, Amino Acid-Specific/*genetics Support, Non-U.S. Gov't Trans-Activators/chemistry/*metabolism Xenopus *Zinc Fingers, Small Nuclear/*genetics RNA, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
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.
