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Blockage of oncostatin M-induced LDL receptor gene transcription by a dominant-negative mutant of C/EBPβ

Zhou, Yue, Zhang, Fang, Abidi, Parveen, Lin, Meihong, Thiel, Gerald, Liu, Jingwen·2006·Biochemical Journal
DOI 10.1042/bj20060044
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Abstract

OM (oncostatin M) activates the human LDLR [LDL (low-density lipoprotein) receptor] gene transcription in HepG2 cells through the SIRE (sterol-independent regulatory element) of LDLR promoter. The SIRE sequence consists of a C/EBP (CCAAT/enhancer-binding protein)-binding site and a CRE (cAMP-response element). Our previous studies [Zhang, Ahlborn, Li, Kraemer and Liu (2002) J. Lipid Res. 43, 1477–1485; Zhang, Lin, Abidi, Thiel and Liu (2003) J. Biol. Chem. 278, 44246–44254] have demonstrated that OM transiently induces EGR-1 (early growth response gene product 1) expression and EGR-1 activates LDLR transcription primarily through a protein–protein interaction with C/EBPβ, which serves as a co-activator of EGR-1. In the present study, we examined the direct role of C/EBPβ as a transactivator in OM-regulated LDLR gene transcription independent of EGR-1. We show that OM induces C/EBPβ expression with kinetics slower than EGR-1 induction. A significant increase in C/EBPβ protein level is detected by 2 h of OM treatment and remains elevated for 24 h. Chromatin immunoprecipitation assays demonstrate that the amount of C/EBPβ bound to the LDLR SIRE sequence is increased 2.8-fold of control by 2 h of OM treatment, reached the highest level of 8-fold by 4 h, and slowly declined thereafter. To further examine the requirement of C/EBPβ in OM-stimulated LDLR expression, we developed a His-tagged dominant-negative mutant of C/EBPβ (His–C/EBPβ-P4; where P4 is plasmid 4 in our mutation series), consisting of the DNA-binding and leucine zipper domains of C/EBPβ (amino acids 246–345). Expression of His–C/EBPβ-P4 in HepG2 cells significantly diminishes the OM-induced increase in LDLR promoter activity and the elevation of endogenous LDLR mRNA expression. Taken together, these new findings identify C/EBPβ as an OM-induced transactivator in LDLR gene transcription and provide a better understanding of the molecular mechanism underlying the sterol-independent regulation of LDLR expression.

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Research works printed but not linked: 24

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Metadata

Authors
Zhou, Y., Zhang, F., Abidi, P., Lin, M., Thiel, G., & Liu, J.
Affiliations
  • Department of Veterans Affairs Palo Alto Health Care System, 3801 Miranda Avenue, Palo Alto, CA 94304, U.S.A.
  • University of Saarland Medical Center, Homburg, Germany
Venue
Biochemical Journal
Publisher
Portland Press Ltd.
Published
2006-06-14
Volume
397
Issue
1
Pages
101-108
ISSN
  • 0264-6021
  • 1470-8728
DOI
10.1042/bj20060044
Cited by
8
References
2
Language
en

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