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Article: The proximal cis-acting elements Sp1, Sp3 and E2F regulate mouse mer gene transcription in sertoli cells

TitleThe proximal cis-acting elements Sp1, Sp3 and E2F regulate mouse mer gene transcription in sertoli cells
Authors
KeywordsE2F
Mer gene
Receptor tyrosine kinases
Sertoli cells
Sp1 and Sp3
Issue Date2002
PublisherBlackwell Publishing Ltd. The Journal's web site is located at http://www.blackwellpublishing.com/journals/EJB
Citation
European Journal Of Biochemistry, 2002, v. 269 n. 15, p. 3789-3800 How to Cite?
AbstractMer belongs to the Tyro 3 family of receptor tyrosine kinases (RTKs). Together with Axl and Rse, the three RTKs are believed to play important functional roles in the male gonads because gene knockout male mice lacking all of these receptors are infertile. In the present study, postnatal expression of Axl and Rse in mouse testes decreased during maturation while expression of Mer increased age-dependently during testicular development. To investigate the transcriptional regulation of gene expression in the testis, a ≈ 1.5 kb fragment of the 5′ flanking sequence of Mer was isolated. The sequence lacks a typical TATA or CAAT box. 5′ RACE revealed that the putative major transcriptional start site of Mer is located at +102 bp upstream of the translation initiation site. Using transient transfections of luciferase reporter constructs driven by various lengths of the 5′ flanking sequence, the gene segment -321/+126 showed the highest transcriptional activity in a mouse Sertoli cell line (TM4). DNAase I footprinting experiments revealed four footprints within the region from -321 to -26, including three binding sites for the transcriptional factor Specificity protein 1 (Sp1) and one for an unknown transcriptional factor. Electrophoretic mobility shift assay (EMSA), supershift assay, mutation studies and cotransfection demonstrated that those Sp1 cis-acting motifs interacted either with Sp1 or Sp1/Sp3, depending on location and the nearby nucleotide sequences. An E2F binding site which down-regulates Mer transcription, as revealed by EMSA, deletion and mutation studies, was identified downstream in the proximity of the promoter. Taking all of these data together, the study has demonstrated that Sp1, Sp3, E2F and probably another unknown transcriptional factor play a critical role in regulating the proximal promoter activities of Mer.
Persistent Identifierhttp://hdl.handle.net/10722/48686
ISSN
2006 Impact Factor: 3.579
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorWong, CCSen_HK
dc.contributor.authorLee, WMen_HK
dc.date.accessioned2008-05-22T04:21:22Z-
dc.date.available2008-05-22T04:21:22Z-
dc.date.issued2002en_HK
dc.identifier.citationEuropean Journal Of Biochemistry, 2002, v. 269 n. 15, p. 3789-3800en_HK
dc.identifier.issn0014-2956en_HK
dc.identifier.urihttp://hdl.handle.net/10722/48686-
dc.description.abstractMer belongs to the Tyro 3 family of receptor tyrosine kinases (RTKs). Together with Axl and Rse, the three RTKs are believed to play important functional roles in the male gonads because gene knockout male mice lacking all of these receptors are infertile. In the present study, postnatal expression of Axl and Rse in mouse testes decreased during maturation while expression of Mer increased age-dependently during testicular development. To investigate the transcriptional regulation of gene expression in the testis, a ≈ 1.5 kb fragment of the 5′ flanking sequence of Mer was isolated. The sequence lacks a typical TATA or CAAT box. 5′ RACE revealed that the putative major transcriptional start site of Mer is located at +102 bp upstream of the translation initiation site. Using transient transfections of luciferase reporter constructs driven by various lengths of the 5′ flanking sequence, the gene segment -321/+126 showed the highest transcriptional activity in a mouse Sertoli cell line (TM4). DNAase I footprinting experiments revealed four footprints within the region from -321 to -26, including three binding sites for the transcriptional factor Specificity protein 1 (Sp1) and one for an unknown transcriptional factor. Electrophoretic mobility shift assay (EMSA), supershift assay, mutation studies and cotransfection demonstrated that those Sp1 cis-acting motifs interacted either with Sp1 or Sp1/Sp3, depending on location and the nearby nucleotide sequences. An E2F binding site which down-regulates Mer transcription, as revealed by EMSA, deletion and mutation studies, was identified downstream in the proximity of the promoter. Taking all of these data together, the study has demonstrated that Sp1, Sp3, E2F and probably another unknown transcriptional factor play a critical role in regulating the proximal promoter activities of Mer.en_HK
dc.format.extent456154 bytes-
dc.format.extent2457 bytes-
dc.format.mimetypeapplication/pdf-
dc.format.mimetypetext/plain-
dc.languageengen_HK
dc.publisherBlackwell Publishing Ltd. The Journal's web site is located at http://www.blackwellpublishing.com/journals/EJBen_HK
dc.relation.ispartofEuropean Journal of Biochemistryen_HK
dc.rightsEuropean Journal of Biochemistry. Copyright © Blackwell Publishing Ltd.en_HK
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.rightsThe definitive version is available at www.blackwell-synergy.comen_HK
dc.subjectE2Fen_HK
dc.subjectMer geneen_HK
dc.subjectReceptor tyrosine kinasesen_HK
dc.subjectSertoli cellsen_HK
dc.subjectSp1 and Sp3en_HK
dc.titleThe proximal cis-acting elements Sp1, Sp3 and E2F regulate mouse mer gene transcription in sertoli cellsen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0014-2956&volume=269&issue=15&spage=3789&epage=3800&date=2002&atitle=The+proximal+cis-acting+elements+Sp1,+Sp3+and+E2F+regulate+mouse+mer+gene+transcription+in+Sertoli+cellsen_HK
dc.identifier.emailLee, WM: hrszlwm@hku.hken_HK
dc.identifier.authorityLee, WM=rp00728en_HK
dc.description.naturepostprinten_HK
dc.identifier.doi10.1046/j.1432-1033.2002.03092.xen_HK
dc.identifier.pmid12153576en_HK
dc.identifier.scopuseid_2-s2.0-0036370524en_HK
dc.identifier.hkuros69752-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0036370524&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume269en_HK
dc.identifier.issue15en_HK
dc.identifier.spage3789en_HK
dc.identifier.epage3800en_HK
dc.identifier.isiWOS:000177782100020-
dc.publisher.placeUnited Kingdomen_HK
dc.identifier.scopusauthoridWong, CCS=19037067100en_HK
dc.identifier.scopusauthoridLee, WM=24799156600en_HK

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