File Download

There are no files associated with this item.

  Links for fulltext
     (May Require Subscription)
Supplementary

Article: The TATA-containing core promoter of the type II collagen gene (COL2A1) is the target of interferon-γ-mediated inhibition in human chondrocytes: Requirement for Stat1α, Jak1 and Jak2

TitleThe TATA-containing core promoter of the type II collagen gene (COL2A1) is the target of interferon-γ-mediated inhibition in human chondrocytes: Requirement for Stat1α, Jak1 and Jak2
Authors
Issue Date2003
PublisherPortland Press Ltd. The Journal's web site is located at http://www.biochemj.org
Citation
Biochemical Journal, 2003, v. 369 n. 1, p. 103-115 How to Cite?
AbstractInterferon-γ (IFN-γ) inhibits the synthesis of the cartilage-specific extracellular matrix protein type II collagen, and suppresses the expression of the type II collagen gene (COL2A1) at the transcriptional level. To further examine this mechanism, the responses of COL2A1 regulatory sequences to IFN-γ and the role of components of the Janus kinase/signal transducer and activators of transcription (JAK/STAT) pathway were examined in the immortalized human chondrocyte cell line, C-28/I2. IFN-γ inhibited the mRNA levels of COL2A1 and aggrecan, but not Sox9, L-Sox5 and Sox6, all of which were expressed by these cells as markers of the differentiated phenotype. IFN-γ suppressed the expression of luciferase reporter constructs containing sequences of the COL2A1 promoter spanning -6368 to +125 bp in the absence and presence of the intronic enhancer and stimulated activity of the γ-interferon-activated site (GAS) luciferase reporter vector, associated with induction of Stat1α-binding activity in nuclear extracts. These responses to IFN-γ were blocked by overexpression of the JAK inhibitor, JAK-binding protein (JAB), or reversed by dominant-negative Stat1α Y701F containing a mutation at Tyr-701, the JAK phosphorylation site. IFN-γ had no effect on COL2A1 promoter expression in Jak1 (U4A)-, Jak2 (γ2A)- and Stat1α (U3A)-deficient cell lines. In the U3A cell line, the response to IFN-γ was rescued by overexpression of Stat1α, but not by either Stat1α Y701F or Stat1β. Functional analysis using deletion constructs showed that the IFN-γ response was retained in the COL2A1 core promoter region spanning -45 to +11 bp, containing the TATA-box and GC-rich sequences but no Stat1-binding elements. Inhibition of COL2A1 promoter activity by IFN-γ persisted in the presence of multiple deletions within the -45/+11 bp region. Our results indicate that repression of COL2A1 gene transcription by IFN-γ requires Jak1, Jak2 and Stat1α and suggest that this response involves indirect interaction of activated Stat1α with the general transcriptional machinery that drives constitutive COL2A1 expression.
Persistent Identifierhttp://hdl.handle.net/10722/147478
ISSN
2015 Impact Factor: 3.562
2015 SCImago Journal Rankings: 2.582
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorOsaki, Men_US
dc.contributor.authorTan, Len_US
dc.contributor.authorChoy, BKen_US
dc.contributor.authorYoshida, Yen_US
dc.contributor.authorCheah, KSEen_US
dc.contributor.authorAuron, PEen_US
dc.contributor.authorGoldring, MBen_US
dc.date.accessioned2012-05-29T06:04:00Z-
dc.date.available2012-05-29T06:04:00Z-
dc.date.issued2003en_US
dc.identifier.citationBiochemical Journal, 2003, v. 369 n. 1, p. 103-115en_US
dc.identifier.issn0264-6021en_US
dc.identifier.urihttp://hdl.handle.net/10722/147478-
dc.description.abstractInterferon-γ (IFN-γ) inhibits the synthesis of the cartilage-specific extracellular matrix protein type II collagen, and suppresses the expression of the type II collagen gene (COL2A1) at the transcriptional level. To further examine this mechanism, the responses of COL2A1 regulatory sequences to IFN-γ and the role of components of the Janus kinase/signal transducer and activators of transcription (JAK/STAT) pathway were examined in the immortalized human chondrocyte cell line, C-28/I2. IFN-γ inhibited the mRNA levels of COL2A1 and aggrecan, but not Sox9, L-Sox5 and Sox6, all of which were expressed by these cells as markers of the differentiated phenotype. IFN-γ suppressed the expression of luciferase reporter constructs containing sequences of the COL2A1 promoter spanning -6368 to +125 bp in the absence and presence of the intronic enhancer and stimulated activity of the γ-interferon-activated site (GAS) luciferase reporter vector, associated with induction of Stat1α-binding activity in nuclear extracts. These responses to IFN-γ were blocked by overexpression of the JAK inhibitor, JAK-binding protein (JAB), or reversed by dominant-negative Stat1α Y701F containing a mutation at Tyr-701, the JAK phosphorylation site. IFN-γ had no effect on COL2A1 promoter expression in Jak1 (U4A)-, Jak2 (γ2A)- and Stat1α (U3A)-deficient cell lines. In the U3A cell line, the response to IFN-γ was rescued by overexpression of Stat1α, but not by either Stat1α Y701F or Stat1β. Functional analysis using deletion constructs showed that the IFN-γ response was retained in the COL2A1 core promoter region spanning -45 to +11 bp, containing the TATA-box and GC-rich sequences but no Stat1-binding elements. Inhibition of COL2A1 promoter activity by IFN-γ persisted in the presence of multiple deletions within the -45/+11 bp region. Our results indicate that repression of COL2A1 gene transcription by IFN-γ requires Jak1, Jak2 and Stat1α and suggest that this response involves indirect interaction of activated Stat1α with the general transcriptional machinery that drives constitutive COL2A1 expression.en_US
dc.languageengen_US
dc.publisherPortland Press Ltd. The Journal's web site is located at http://www.biochemj.orgen_US
dc.relation.ispartofBiochemical Journalen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshCell Line, Transformeden_US
dc.subject.meshChondrocytes - Metabolismen_US
dc.subject.meshCollagen Type Ii - Geneticsen_US
dc.subject.meshDna - Metabolismen_US
dc.subject.meshHumansen_US
dc.subject.meshInterferon-Stimulated Gene Factor 3en_US
dc.subject.meshInterferon-Gamma - Physiologyen_US
dc.subject.meshJanus Kinase 1en_US
dc.subject.meshJanus Kinase 2en_US
dc.subject.meshMolecular Sequence Dataen_US
dc.subject.meshMutagenesis, Site-Directeden_US
dc.subject.meshPromoter Regions, Geneticen_US
dc.subject.meshProtein-Tyrosine Kinases - Metabolismen_US
dc.subject.meshProto-Oncogene Proteinsen_US
dc.subject.meshRna, Messenger - Geneticsen_US
dc.subject.meshSequence Homology, Nucleic Aciden_US
dc.subject.meshTata Boxen_US
dc.subject.meshTranscription Factors - Metabolismen_US
dc.titleThe TATA-containing core promoter of the type II collagen gene (COL2A1) is the target of interferon-γ-mediated inhibition in human chondrocytes: Requirement for Stat1α, Jak1 and Jak2en_US
dc.typeArticleen_US
dc.identifier.emailCheah, KSE:hrmbdkc@hku.hken_US
dc.identifier.authorityCheah, KSE=rp00342en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1042/BJ20020928en_US
dc.identifier.pmid12223098en_US
dc.identifier.scopuseid_2-s2.0-0037269828en_US
dc.identifier.hkuros80347-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0037269828&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume369en_US
dc.identifier.issue1en_US
dc.identifier.spage103en_US
dc.identifier.epage115en_US
dc.identifier.isiWOS:000180871000012-
dc.publisher.placeUnited Kingdomen_US
dc.identifier.scopusauthoridOsaki, M=55198849200en_US
dc.identifier.scopusauthoridTan, L=7402233616en_US
dc.identifier.scopusauthoridChoy, BK=7003465497en_US
dc.identifier.scopusauthoridYoshida, Y=7404399408en_US
dc.identifier.scopusauthoridCheah, KSE=35387746200en_US
dc.identifier.scopusauthoridAuron, PE=7006635911en_US
dc.identifier.scopusauthoridGoldring, MB=7005248020en_US

Export via OAI-PMH Interface in XML Formats


OR


Export to Other Non-XML Formats