New mode of DNA binding of multi-zinc finger transcription factors: delta EF1 family members bind with two hands to two target sites

Autor: Harry Kraft, Clara Collart, Gunther Wuytens, Walter Lerchner, Jacques E. Remacle, Danny Huylebroeck, Kristin Verschueren, James C. Smith
Rok vydání: 1999
Předmět:
Integrin alpha4
Molecular Sequence Data
Gene Expression
Repressor
In Vitro Techniques
Xenopus Proteins
Biology
DNA-binding protein
General Biochemistry
Genetics and Molecular Biology
Animals
Genetically Modified
Xenopus laevis
Antigens
CD
Genes
Reporter
Animals
Humans
EF Hand Motifs
Binding site
Promoter Regions
Genetic
Molecular Biology
Gene
Zinc Finger E-box Binding Homeobox 2
Homeodomain Proteins
Zinc finger
Genetics
Binding Sites
Base Sequence
General Immunology and Microbiology
General Neuroscience
Nuclear Proteins
Zinc Finger E-box-Binding Homeobox 1
Zinc Fingers
DNA
Cadherins
Zinc finger nuclease
Cell biology
DNA-Binding Proteins
Repressor Proteins
DNA binding site
COS Cells
Mutation
Female
DNA Probes
T-Box Domain Proteins
Research Article
Protein Binding
Transcription Factors
Binding domain
Zdroj: The EMBO Journal. 18:5073-5084
ISSN: 1460-2075
Popis: SIP1, a Smad-interacting protein, and deltaEF1, a transcriptional repressor involved in skeletal and T-cell development, belong to the same family of DNA binding proteins. SIP1 and deltaEF1 contain two separated clusters of zinc fingers, one N-terminal and one C-terminal. These clusters show high sequence homology and are highly conserved between SIP1 and deltaEF1. Each zinc finger cluster binds independently to a 5'-CACCT sequence. However, high-affinity binding sites for full-length SIP1 and deltaEF1 in the promoter regions of candidate target genes like Xenopus Xbra2, and human alpha4-integrin and E-cadherin, are bipartite elements composed of one CACCT and one CACCTG sequence, the orientation and spacing of which can vary. Using transgenic Xenopus embryos, we demonstrate that the integrity of these two sequences is necessary for correct spatial expression of a Xbra2 promoter-driven reporter gene. Both zinc finger clusters must be intact for the high-affinity binding of SIP1 to DNA and for its optimal repressor activity. Our results show that SIP1 binds as monomer and contacts one target sequence with the first zinc finger cluster, and the other with the second cluster. Our work redefines the optimal binding site and, consequently, candidate target genes for vertebrate members of the deltaEF1 family.
Databáze: OpenAIRE
Externí odkaz: