Hsp90α recruited by Sp1 is important for transcription of 12(S)-lipoxygenase in A431 cells

Jan Jong Hung, Chih Ying Wu, Pao Chi Liao, Wen Chang Chang

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Sp1 is a basic transcriptional factor that binds to the GC-rich region in the promoter of the target gene. It is involved in transcription of numerous genes by recruiting transcriptional factors to the promoters of target genes. In this study, we found in vivo and in vitro that Hsp90α was recruited to the GC-rich region of the 12(S)-lipoxygenase promoter through interaction with Sp1 in A431 cells by employing DNA affinity immunoprecipitation assay and chromatin immunoprecipitation assay. When Hsp90α was inhibited by geldanamycin (GA, a specific inhibitor of the Hsp90 family) or by siRNA of Hsp90α (to block its activity or to knockdown protein levels), respectively, luciferase activity (driven by the 12(S)-lipoxygenase promoter) and both mRNA and protein levels of 12(S)-lipoxygenase were reduced significantly in cells. In addition, the effect of GA was abolished when the Sp1 binding sites of 12(S)-lipoxygenase were mutated in A431 cells. Interestingly, binding of Sp1 to the 12(S)-lipoxygenase promoter was also decreased upon GA treatment in cells. In conclusion, our results indicate that Sp1 interacts with Hsp90α to recruit it to the promoter of 12(S)-lipoxygenase and then to regulate gene transcription by modulating the binding ability of Sp1 to promoters.

Original languageEnglish
Pages (from-to)36283-36292
Number of pages10
JournalJournal of Biological Chemistry
Volume280
Issue number43
DOIs
Publication statusPublished - Oct 28 2005
Externally publishedYes

Fingerprint

Arachidonate 12-Lipoxygenase
Lipoxygenase
Transcription
Genes
GC Rich Sequence
Assays
Chromatin Immunoprecipitation
Luciferases
Immunoprecipitation
Small Interfering RNA
Chromatin
Proteins
Binding Sites
Messenger RNA
DNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

Hsp90α recruited by Sp1 is important for transcription of 12(S)-lipoxygenase in A431 cells. / Hung, Jan Jong; Wu, Chih Ying; Liao, Pao Chi; Chang, Wen Chang.

In: Journal of Biological Chemistry, Vol. 280, No. 43, 28.10.2005, p. 36283-36292.

Research output: Contribution to journalArticle

Hung, Jan Jong ; Wu, Chih Ying ; Liao, Pao Chi ; Chang, Wen Chang. / Hsp90α recruited by Sp1 is important for transcription of 12(S)-lipoxygenase in A431 cells. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 43. pp. 36283-36292.
@article{3a9f603d820844a9918895fde7780a2d,
title = "Hsp90α recruited by Sp1 is important for transcription of 12(S)-lipoxygenase in A431 cells",
abstract = "Sp1 is a basic transcriptional factor that binds to the GC-rich region in the promoter of the target gene. It is involved in transcription of numerous genes by recruiting transcriptional factors to the promoters of target genes. In this study, we found in vivo and in vitro that Hsp90α was recruited to the GC-rich region of the 12(S)-lipoxygenase promoter through interaction with Sp1 in A431 cells by employing DNA affinity immunoprecipitation assay and chromatin immunoprecipitation assay. When Hsp90α was inhibited by geldanamycin (GA, a specific inhibitor of the Hsp90 family) or by siRNA of Hsp90α (to block its activity or to knockdown protein levels), respectively, luciferase activity (driven by the 12(S)-lipoxygenase promoter) and both mRNA and protein levels of 12(S)-lipoxygenase were reduced significantly in cells. In addition, the effect of GA was abolished when the Sp1 binding sites of 12(S)-lipoxygenase were mutated in A431 cells. Interestingly, binding of Sp1 to the 12(S)-lipoxygenase promoter was also decreased upon GA treatment in cells. In conclusion, our results indicate that Sp1 interacts with Hsp90α to recruit it to the promoter of 12(S)-lipoxygenase and then to regulate gene transcription by modulating the binding ability of Sp1 to promoters.",
author = "Hung, {Jan Jong} and Wu, {Chih Ying} and Liao, {Pao Chi} and Chang, {Wen Chang}",
year = "2005",
month = "10",
day = "28",
doi = "10.1074/jbc.M504904200",
language = "English",
volume = "280",
pages = "36283--36292",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "43",

}

TY - JOUR

T1 - Hsp90α recruited by Sp1 is important for transcription of 12(S)-lipoxygenase in A431 cells

AU - Hung, Jan Jong

AU - Wu, Chih Ying

AU - Liao, Pao Chi

AU - Chang, Wen Chang

PY - 2005/10/28

Y1 - 2005/10/28

N2 - Sp1 is a basic transcriptional factor that binds to the GC-rich region in the promoter of the target gene. It is involved in transcription of numerous genes by recruiting transcriptional factors to the promoters of target genes. In this study, we found in vivo and in vitro that Hsp90α was recruited to the GC-rich region of the 12(S)-lipoxygenase promoter through interaction with Sp1 in A431 cells by employing DNA affinity immunoprecipitation assay and chromatin immunoprecipitation assay. When Hsp90α was inhibited by geldanamycin (GA, a specific inhibitor of the Hsp90 family) or by siRNA of Hsp90α (to block its activity or to knockdown protein levels), respectively, luciferase activity (driven by the 12(S)-lipoxygenase promoter) and both mRNA and protein levels of 12(S)-lipoxygenase were reduced significantly in cells. In addition, the effect of GA was abolished when the Sp1 binding sites of 12(S)-lipoxygenase were mutated in A431 cells. Interestingly, binding of Sp1 to the 12(S)-lipoxygenase promoter was also decreased upon GA treatment in cells. In conclusion, our results indicate that Sp1 interacts with Hsp90α to recruit it to the promoter of 12(S)-lipoxygenase and then to regulate gene transcription by modulating the binding ability of Sp1 to promoters.

AB - Sp1 is a basic transcriptional factor that binds to the GC-rich region in the promoter of the target gene. It is involved in transcription of numerous genes by recruiting transcriptional factors to the promoters of target genes. In this study, we found in vivo and in vitro that Hsp90α was recruited to the GC-rich region of the 12(S)-lipoxygenase promoter through interaction with Sp1 in A431 cells by employing DNA affinity immunoprecipitation assay and chromatin immunoprecipitation assay. When Hsp90α was inhibited by geldanamycin (GA, a specific inhibitor of the Hsp90 family) or by siRNA of Hsp90α (to block its activity or to knockdown protein levels), respectively, luciferase activity (driven by the 12(S)-lipoxygenase promoter) and both mRNA and protein levels of 12(S)-lipoxygenase were reduced significantly in cells. In addition, the effect of GA was abolished when the Sp1 binding sites of 12(S)-lipoxygenase were mutated in A431 cells. Interestingly, binding of Sp1 to the 12(S)-lipoxygenase promoter was also decreased upon GA treatment in cells. In conclusion, our results indicate that Sp1 interacts with Hsp90α to recruit it to the promoter of 12(S)-lipoxygenase and then to regulate gene transcription by modulating the binding ability of Sp1 to promoters.

UR - http://www.scopus.com/inward/record.url?scp=27744586073&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=27744586073&partnerID=8YFLogxK

U2 - 10.1074/jbc.M504904200

DO - 10.1074/jbc.M504904200

M3 - Article

VL - 280

SP - 36283

EP - 36292

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 43

ER -