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Article: Sp1 site is crucial for the mouse claudin-19 gene expression in the kidney cells

TitleSp1 site is crucial for the mouse claudin-19 gene expression in the kidney cells
Authors
Issue Date2004
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/febslet
Citation
Febs Letters, 2004, v. 578 n. 3, p. 251-256 How to Cite?
AbstractMembers of the claudin family play important roles in the formation of tight junctions in the kidneys, liver and intestine. Claudin-19 (Cldn19), a newly identified member of this family, is highly expressed in the kidney of the mouse. To have a better understanding on mouse claudin-19 gene expression, a 0.9-kb DNA fragment containing the 5′-flanking region of the Cldn19 gene was isolated. DNA sequence comparison between the mouse and human Cldn19 promoter regions exhibited little homology. One transcription initiation site was located at 104 nucleotides upstream of the start codon (ATG) of the Cldn19 gene. The mouse claudin-19 promoter lacked typical CAAT or GC-box. Deletion constructs of the 0.9-kb DNA fragment were generated and fused to a promoterless luciferase (Luc) reporter plasmid. Transfection studies using various kidney cell lines (MDCK, mIMCD3 and HEK293) revealed that the minimal promoter fragment resided in the -39 to -108 region, which contained a number of binding sites for transcription factors including Sp1. Site-directed mutagenesis using specific oligo probes confirmed that Sp1 was crucial for Cldn19 transactivation in the three cell lines studied. Electromobility shift assay confirmed that the nuclear extracts of these cells bound to the Sp1 oligo derived from Cldn19 promoter, but not to the mutated Sp1 oligo probe. Moreover, this DNA-protein complex would be recognized by Sp1 antibody, indicating specific Sp1 binding. Collectively, our data suggest that Sp1 binds to the claudin-19 promoter region and is responsible for its expression in the kidney cell lines in vitro. © 2004 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.
Persistent Identifierhttp://hdl.handle.net/10722/173273
ISSN
2015 Impact Factor: 3.519
2015 SCImago Journal Rankings: 2.026
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorLuk, JMen_US
dc.contributor.authorTong, MKen_US
dc.contributor.authorMok, BWen_US
dc.contributor.authorTam, PCen_US
dc.contributor.authorYeung, WSBen_US
dc.contributor.authorLee, KFen_US
dc.date.accessioned2012-10-30T06:28:58Z-
dc.date.available2012-10-30T06:28:58Z-
dc.date.issued2004en_US
dc.identifier.citationFebs Letters, 2004, v. 578 n. 3, p. 251-256en_US
dc.identifier.issn0014-5793en_US
dc.identifier.urihttp://hdl.handle.net/10722/173273-
dc.description.abstractMembers of the claudin family play important roles in the formation of tight junctions in the kidneys, liver and intestine. Claudin-19 (Cldn19), a newly identified member of this family, is highly expressed in the kidney of the mouse. To have a better understanding on mouse claudin-19 gene expression, a 0.9-kb DNA fragment containing the 5′-flanking region of the Cldn19 gene was isolated. DNA sequence comparison between the mouse and human Cldn19 promoter regions exhibited little homology. One transcription initiation site was located at 104 nucleotides upstream of the start codon (ATG) of the Cldn19 gene. The mouse claudin-19 promoter lacked typical CAAT or GC-box. Deletion constructs of the 0.9-kb DNA fragment were generated and fused to a promoterless luciferase (Luc) reporter plasmid. Transfection studies using various kidney cell lines (MDCK, mIMCD3 and HEK293) revealed that the minimal promoter fragment resided in the -39 to -108 region, which contained a number of binding sites for transcription factors including Sp1. Site-directed mutagenesis using specific oligo probes confirmed that Sp1 was crucial for Cldn19 transactivation in the three cell lines studied. Electromobility shift assay confirmed that the nuclear extracts of these cells bound to the Sp1 oligo derived from Cldn19 promoter, but not to the mutated Sp1 oligo probe. Moreover, this DNA-protein complex would be recognized by Sp1 antibody, indicating specific Sp1 binding. Collectively, our data suggest that Sp1 binds to the claudin-19 promoter region and is responsible for its expression in the kidney cell lines in vitro. © 2004 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.en_US
dc.languageengen_US
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/febsleten_US
dc.relation.ispartofFEBS Lettersen_US
dc.rightsF E B S Letters. Copyright © Elsevier BV.-
dc.subject.mesh5' Flanking Regionen_US
dc.subject.meshAmino Acid Sequenceen_US
dc.subject.meshAnimalsen_US
dc.subject.meshAntibodies, Monoclonal - Metabolismen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshBinding Sitesen_US
dc.subject.meshCell Lineen_US
dc.subject.meshCodon, Initiatoren_US
dc.subject.meshDogsen_US
dc.subject.meshElectrophoretic Mobility Shift Assayen_US
dc.subject.meshExonsen_US
dc.subject.meshGene Expression Regulationen_US
dc.subject.meshGenes, Reporteren_US
dc.subject.meshHumansen_US
dc.subject.meshKidney - Cytologyen_US
dc.subject.meshLuciferases - Metabolismen_US
dc.subject.meshMembrane Proteins - Genetics - Metabolismen_US
dc.subject.meshMiceen_US
dc.subject.meshMolecular Sequence Dataen_US
dc.subject.meshMutagenesis, Site-Directeden_US
dc.subject.meshPromoter Regions, Geneticen_US
dc.subject.meshProtein Bindingen_US
dc.subject.meshSp1 Transcription Factor - Genetics - Metabolismen_US
dc.subject.meshTranscription Initiation Siteen_US
dc.subject.meshTranscriptional Activationen_US
dc.titleSp1 site is crucial for the mouse claudin-19 gene expression in the kidney cellsen_US
dc.typeArticleen_US
dc.identifier.emailLuk, JM:jmluk@hkucc.hku.hken_US
dc.identifier.emailYeung, WSB:wsbyeung@hkucc.hku.hken_US
dc.identifier.emailLee, KF:ckflee@hku.hken_US
dc.identifier.authorityLuk, JM=rp00349en_US
dc.identifier.authorityYeung, WSB=rp00331en_US
dc.identifier.authorityLee, KF=rp00458en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1016/j.febslet.2004.11.010en_US
dc.identifier.pmid15589828-
dc.identifier.scopuseid_2-s2.0-10044248760en_US
dc.identifier.hkuros97179-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-10044248760&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume578en_US
dc.identifier.issue3en_US
dc.identifier.spage251en_US
dc.identifier.epage256en_US
dc.identifier.isiWOS:000225939900010-
dc.publisher.placeNetherlandsen_US
dc.identifier.scopusauthoridLuk, JM=7006777791en_US
dc.identifier.scopusauthoridTong, MK=36880476300en_US
dc.identifier.scopusauthoridMok, BW=36786012900en_US
dc.identifier.scopusauthoridTam, PC=7202539419en_US
dc.identifier.scopusauthoridYeung, WSB=7102370745en_US
dc.identifier.scopusauthoridLee, KF=26643097500en_US

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