Cux/CDP homeodomain protein binds to an enhancer in the rat c- mos locus and represses its activity
Identifieur interne : 003C78 ( Main/Exploration ); précédent : 003C77; suivant : 003C79Cux/CDP homeodomain protein binds to an enhancer in the rat c- mos locus and represses its activity
Auteurs : Nadia A. Higgy [Canada] ; Heide A. Tarnasky [Canada] ; Isabelle Valarché [Pays-Bas] ; Alain Nepveu [Canada] ; Frans A. Van Der Hoorn [Canada]Source :
- BBA - Gene Structure and Expression [ 0167-4781 ] ; 1997.
English descriptors
- Teeft :
- Acta, Amino acids, Assay, Bacterial extracts, Binding site, Biochimica, Biol, Biophysica, Biophysica acta, Cdna, Cdp2, Cell cycle, Cuxrcdp, Cuxrcdp proteins, Cuxrcdp repression, Deletion, Deletion mutants, Dnase, Ectopic expression, Emsa, Emsa supershift assays, Enhancer, Enhancer activity, Enhancer site, Fibroblast, Footprint, Fusion protein, Hcut, Higgy, Homeodomain, Homeodomain protein, Hoorn, Internal control, Monoclonal, Monoclonal antibodies, Mouse fibroblasts, Mouse lung, Mouse testis, Murine, Murine cuxrcdp, Murine lung, Mutant, Mutant site, Nepveu, Nuclear extracts, Nuclear factor, Nuclear protein, Nucleic acids, Oligo, Oocyte, Pblcat2, Phage, Plasmid, Prcrcmvcuxfl, Promoter, Reporter plasmid, Repressor, Somatic cells, Testis, Transfection, Transfections, Unrelated control, Vande woude.
Abstract
Abstract: The c-mos gene is transcribed in male and female germ cells, in differentiating myoblasts and in 3T3 cells from cell-specific promoters. We characterized the rat testis promoter, which contains a TATA-box and one binding site for a testis-specific transcription factor TTF-D, as well as a region which can act as enhancer, which is located approx. 2 kb upstream of the c-mos AUG start codon. It binds three factors at sites I, II and III as determined in DNAse I footprint assays. We demonstrated that a member of the NF-1/CTF family of transcription factors binds site II. Here we report the cloning of the protein that binds to enhancer site III. This protein is the rat homolog of human hCut/CDP, mouse Cux/CDP and canine Clox. hCut/Cux/CDP/Clox (hereafter called Cux/CDP), a 160 kDa protein containing multiple repeats and a homeodomain, negatively regulates the mammalian c-myc, gp91-phox and N-Cam genes. Using bacterially produced murine GST-Cux fusion proteins and GST-Cux deletion mutants, we find that Cux repeat CR3 and the homeodomain are both required for efficient binding to enhancer site III. Mouse lung and testis nuclear Cux/CDP bind to site III as determined in electrophoretic gel mobility supershift assays using two different anti-hCut specific monoclonal antibodies. Transfections of CAT constructs containing the enhancer fragment linked to a minimal promoter demonstrated that Cux/CDP represses c-mos enhancer activity.
Url:
DOI: 10.1016/S0167-4781(96)00221-7
Affiliations:
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Le document en format XML
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Van Der Hoorn</name><affiliation wicri:level="4"><country>Canada</country><wicri:regionArea>Department of Medical Biochemistry, University of Calgary, 3330 Hospital Drive, N.W., Calgary, Alberta</wicri:regionArea><orgName type="university">Université de Calgary</orgName><placeName><settlement type="city">Calgary</settlement><region type="state">Alberta</region></placeName></affiliation></author></analytic><monogr></monogr><series><title level="j">BBA - Gene Structure and Expression</title><title level="j" type="abbrev">BBAEXP</title><idno type="ISSN">0167-4781</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1997">1997</date><biblScope unit="volume">1351</biblScope><biblScope unit="issue">3</biblScope><biblScope unit="page" from="313">313</biblScope><biblScope unit="page" to="324">324</biblScope></imprint><idno type="ISSN">0167-4781</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0167-4781</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Acta</term><term>Amino acids</term><term>Assay</term><term>Bacterial extracts</term><term>Binding site</term><term>Biochimica</term><term>Biol</term><term>Biophysica</term><term>Biophysica acta</term><term>Cdna</term><term>Cdp2</term><term>Cell cycle</term><term>Cuxrcdp</term><term>Cuxrcdp proteins</term><term>Cuxrcdp repression</term><term>Deletion</term><term>Deletion mutants</term><term>Dnase</term><term>Ectopic expression</term><term>Emsa</term><term>Emsa supershift assays</term><term>Enhancer</term><term>Enhancer activity</term><term>Enhancer site</term><term>Fibroblast</term><term>Footprint</term><term>Fusion protein</term><term>Hcut</term><term>Higgy</term><term>Homeodomain</term><term>Homeodomain protein</term><term>Hoorn</term><term>Internal control</term><term>Monoclonal</term><term>Monoclonal antibodies</term><term>Mouse fibroblasts</term><term>Mouse lung</term><term>Mouse testis</term><term>Murine</term><term>Murine cuxrcdp</term><term>Murine lung</term><term>Mutant</term><term>Mutant site</term><term>Nepveu</term><term>Nuclear extracts</term><term>Nuclear factor</term><term>Nuclear protein</term><term>Nucleic acids</term><term>Oligo</term><term>Oocyte</term><term>Pblcat2</term><term>Phage</term><term>Plasmid</term><term>Prcrcmvcuxfl</term><term>Promoter</term><term>Reporter plasmid</term><term>Repressor</term><term>Somatic cells</term><term>Testis</term><term>Transfection</term><term>Transfections</term><term>Unrelated control</term><term>Vande woude</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: The c-mos gene is transcribed in male and female germ cells, in differentiating myoblasts and in 3T3 cells from cell-specific promoters. We characterized the rat testis promoter, which contains a TATA-box and one binding site for a testis-specific transcription factor TTF-D, as well as a region which can act as enhancer, which is located approx. 2 kb upstream of the c-mos AUG start codon. It binds three factors at sites I, II and III as determined in DNAse I footprint assays. We demonstrated that a member of the NF-1/CTF family of transcription factors binds site II. Here we report the cloning of the protein that binds to enhancer site III. This protein is the rat homolog of human hCut/CDP, mouse Cux/CDP and canine Clox. hCut/Cux/CDP/Clox (hereafter called Cux/CDP), a 160 kDa protein containing multiple repeats and a homeodomain, negatively regulates the mammalian c-myc, gp91-phox and N-Cam genes. Using bacterially produced murine GST-Cux fusion proteins and GST-Cux deletion mutants, we find that Cux repeat CR3 and the homeodomain are both required for efficient binding to enhancer site III. Mouse lung and testis nuclear Cux/CDP bind to site III as determined in electrophoretic gel mobility supershift assays using two different anti-hCut specific monoclonal antibodies. Transfections of CAT constructs containing the enhancer fragment linked to a minimal promoter demonstrated that Cux/CDP represses c-mos enhancer activity.</div></front></TEI><affiliations><list><country><li>Canada</li><li>Pays-Bas</li></country><region><li>Alberta</li></region><settlement><li>Calgary</li></settlement><orgName><li>Université de Calgary</li></orgName></list><tree><country name="Canada"><region name="Alberta"><name sortKey="Higgy, Nadia A" sort="Higgy, Nadia A" uniqKey="Higgy N" first="Nadia A" last="Higgy">Nadia A. Higgy</name></region><name sortKey="Nepveu, Alain" sort="Nepveu, Alain" uniqKey="Nepveu A" first="Alain" last="Nepveu">Alain Nepveu</name><name sortKey="Tarnasky, Heide A" sort="Tarnasky, Heide A" uniqKey="Tarnasky H" first="Heide A" last="Tarnasky">Heide A. Tarnasky</name><name sortKey="Van Der Hoorn, Frans A" sort="Van Der Hoorn, Frans A" uniqKey="Van Der Hoorn F" first="Frans A" last="Van Der Hoorn">Frans A. Van Der Hoorn</name></country><country name="Pays-Bas"><noRegion><name sortKey="Valarche, Isabelle" sort="Valarche, Isabelle" uniqKey="Valarche I" first="Isabelle" last="Valarché">Isabelle Valarché</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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