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Conference Paper: Transcriptional and post-translational regulations of nectin-like molecule-2 in spermatogenic cells under the control of transforming growth factor-β1

TitleTranscriptional and post-translational regulations of nectin-like molecule-2 in spermatogenic cells under the control of transforming growth factor-β1
Authors
KeywordsBiology
Biochemistry
Issue Date2011
PublisherWiley-Blackwell Publishing Ltd.. The Journal's web site is located at http://www.febsjournal.org/
Citation
The 36th Federation of European Biochemical Societies Congress (FEBS 2011), Torino, Italy, 25-30 June 2011. In The FEBS Journal, 2011, v. 278 suppl. s1, p. 116, abstract P03.83 How to Cite?
AbstractTransforming growth factor-β (TGF-β) is a known family of cytokine that regulates cell junction restructuring required for germ cell development and movement in the testis. Previous studies have shown that it is important to regulate the restructuring of the blood-testis barrier constituted by an array of tight and adherens junctions between adjacent sertoli cells. In this study, we extend our investigation to examine if TGF-β1 regulates the restructuring of cell junctions between sertoli and germ cells. Nectin-like molecule-2 (necl-2) is found in spermatogenic cells and is involved in Sertoli-germ cell interaction, we therefore examine if TGF-β1 exert its effect on Sertoli-germ cell junction restructuring by modulating the expression of necl-2 in spermatogenic cells. In this study, we have found that TGF-β1 (5 ng/ml) significantly downregulates both mRNA and protein levels of necl-2 in mouse germ cell cell line, GC-1 spg cells. The reduction of necl-2 mRNA mediated by TGF-β1 is at transcriptional level, but not post-transcriptional level. Luciferase reporter assays have confirmed that TGF-β1 downregulates necl-2 promoter activity and siRNA analyses have shown that knockdown of Smad3 and Smad4 abolish TGF-β1-mediated necl-2 gene repression. Electromobility shift assays have shown that TGF-β1 activates and promotes the binding of Smad3 and Smad4 to the cis-acting motifs including two CCAAT and MyoD motifs that lie within nt-159 and -1 of the promoter region. Apart from transcriptional regulation, TGF-β1 exerts its negative effect on necl-2 expression via post-translational regulation. Cycloheximide treatment coupled with immunoblotting has shown that TGF-β1 speed-ups the turnover of necl-2 protein. By inhibitor and shRNA approaches targeting targeting clathrin, we have confirmed that TGF-β1 promotes necl-2 degradation by activating clathrin-dependent endocytosis.
DescriptionThis journal suppl. is Special Issue of 36th FEBS 2011 Congress
Poster Presentations: P03 – Protein structure, functional mechanisms, turnover: P03.83
Persistent Identifierhttp://hdl.handle.net/10722/146049
ISSN
2015 Impact Factor: 4.237
2015 SCImago Journal Rankings: 2.141

 

DC FieldValueLanguage
dc.contributor.authorGao, Yen_US
dc.contributor.authorLui, WYen_US
dc.date.accessioned2012-03-27T09:08:27Z-
dc.date.available2012-03-27T09:08:27Z-
dc.date.issued2011en_US
dc.identifier.citationThe 36th Federation of European Biochemical Societies Congress (FEBS 2011), Torino, Italy, 25-30 June 2011. In The FEBS Journal, 2011, v. 278 suppl. s1, p. 116, abstract P03.83en_US
dc.identifier.issn1742-464X-
dc.identifier.urihttp://hdl.handle.net/10722/146049-
dc.descriptionThis journal suppl. is Special Issue of 36th FEBS 2011 Congress-
dc.descriptionPoster Presentations: P03 – Protein structure, functional mechanisms, turnover: P03.83-
dc.description.abstractTransforming growth factor-β (TGF-β) is a known family of cytokine that regulates cell junction restructuring required for germ cell development and movement in the testis. Previous studies have shown that it is important to regulate the restructuring of the blood-testis barrier constituted by an array of tight and adherens junctions between adjacent sertoli cells. In this study, we extend our investigation to examine if TGF-β1 regulates the restructuring of cell junctions between sertoli and germ cells. Nectin-like molecule-2 (necl-2) is found in spermatogenic cells and is involved in Sertoli-germ cell interaction, we therefore examine if TGF-β1 exert its effect on Sertoli-germ cell junction restructuring by modulating the expression of necl-2 in spermatogenic cells. In this study, we have found that TGF-β1 (5 ng/ml) significantly downregulates both mRNA and protein levels of necl-2 in mouse germ cell cell line, GC-1 spg cells. The reduction of necl-2 mRNA mediated by TGF-β1 is at transcriptional level, but not post-transcriptional level. Luciferase reporter assays have confirmed that TGF-β1 downregulates necl-2 promoter activity and siRNA analyses have shown that knockdown of Smad3 and Smad4 abolish TGF-β1-mediated necl-2 gene repression. Electromobility shift assays have shown that TGF-β1 activates and promotes the binding of Smad3 and Smad4 to the cis-acting motifs including two CCAAT and MyoD motifs that lie within nt-159 and -1 of the promoter region. Apart from transcriptional regulation, TGF-β1 exerts its negative effect on necl-2 expression via post-translational regulation. Cycloheximide treatment coupled with immunoblotting has shown that TGF-β1 speed-ups the turnover of necl-2 protein. By inhibitor and shRNA approaches targeting targeting clathrin, we have confirmed that TGF-β1 promotes necl-2 degradation by activating clathrin-dependent endocytosis.-
dc.languageengen_US
dc.publisherWiley-Blackwell Publishing Ltd.. The Journal's web site is located at http://www.febsjournal.org/-
dc.relation.ispartofThe FEBS Journalen_US
dc.rightsThe definitive version is available at www3.interscience.wiley.com-
dc.subjectBiology-
dc.subjectBiochemistry-
dc.titleTranscriptional and post-translational regulations of nectin-like molecule-2 in spermatogenic cells under the control of transforming growth factor-β1en_US
dc.typeConference_Paperen_US
dc.identifier.emailLui, WY: wylui@hku.hken_US
dc.identifier.authorityLui, WY=rp00756en_US
dc.description.naturelink_to_OA_fulltext-
dc.identifier.doi10.1111/j.1742-4658.2011.08137.x-
dc.identifier.hkuros191913en_US
dc.identifier.hkuros198855-
dc.identifier.volume278-
dc.identifier.issuesuppl. s1-
dc.identifier.spage116-
dc.identifier.epage116-
dc.publisher.placeUnited Kingdom-
dc.customcontrol.immutablesml 130327-

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