Identification of the CREB-binding protein/p300-interacting protein CITED2 as a peroxisome proliferator-activated receptor α coregulator

Eric S. Tien, John W. Davis, John P. Vanden Heuvel

Research output: Contribution to journalArticle

90 Citations (Scopus)

Abstract

Like other nuclear receptors, the peroxisome proliferator-activated receptors (PPARs) use a wide variety of protein-protein interactions to properly regulate transcription of target genes. In an attempt to identify novel PPAR-interacting proteins, a cDNA expression library was screened with bacterially expressed PPARα. One of the genes identified as a PPARα-associated protein by interaction cloning was the CREB-binding protein/p300-interacting transactivator with ED-rich tail 2 (CITED2, also called p35srj/mrg1/msg1). This coactivator interacted directly with PPARα in the presence or absence of ligand predominantly via the ligand binding domain of the nuclear receptor. In transient transfection reporter assays, CITED2 acted as a dose-dependent coactivator of PPARα-dependent transcriptional regulation in the presence of several exogenous ligands. CITED2 also increased PPARγ-dependent regulation of reporter genes but had no effect on PPARβ activity. To determine whether CITED2 affects endogenous gene expression, this protein was stably overexpressed (CITED2+) or repressed by small inhibitor RNA (CITED2-) in immortalized mouse hepatocytes. Relative to the control stably transfected or CITED2- cells, CITED2+ cells had an increased rate of cell proliferation. Microarray analysis and real time PCR showed that several genes are differentially affected by PPARα ligands in CITED2+ versus CITED2- cells. Genes that were affected by PPARα ligands in a CITED2-modulatory manner include angiopoietin-like protein 4, forkhead C2, hypoxia-inducible factor-1α, and MAPK phosphatase 1. Interestingly these genes share common functions in that they are known to promote vascularization and angiogenesis in response to hypoxia. The results described here suggest that CIT-ED2 is a coactivator of PPARα and that both proteins may participate in signaling cascades of hypoxic response and angiogenesis.

Original languageEnglish (US)
Pages (from-to)24053-24063
Number of pages11
JournalJournal of Biological Chemistry
Volume279
Issue number23
DOIs
StatePublished - Jun 4 2004

Fingerprint

CREB-Binding Protein
Peroxisome Proliferator-Activated Receptors
Genes
Proteins
Ligands
Cytoplasmic and Nuclear Receptors
Dual Specificity Phosphatase 1
Angiopoietins
Receptor-Interacting Protein Serine-Threonine Kinases
Forkhead Transcription Factors
Hypoxia-Inducible Factor 1
Trans-Activators
Cloning
Cell proliferation
Transcription
Microarray Analysis
Microarrays
Gene Library
Reporter Genes
Gene expression

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

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title = "Identification of the CREB-binding protein/p300-interacting protein CITED2 as a peroxisome proliferator-activated receptor α coregulator",
abstract = "Like other nuclear receptors, the peroxisome proliferator-activated receptors (PPARs) use a wide variety of protein-protein interactions to properly regulate transcription of target genes. In an attempt to identify novel PPAR-interacting proteins, a cDNA expression library was screened with bacterially expressed PPARα. One of the genes identified as a PPARα-associated protein by interaction cloning was the CREB-binding protein/p300-interacting transactivator with ED-rich tail 2 (CITED2, also called p35srj/mrg1/msg1). This coactivator interacted directly with PPARα in the presence or absence of ligand predominantly via the ligand binding domain of the nuclear receptor. In transient transfection reporter assays, CITED2 acted as a dose-dependent coactivator of PPARα-dependent transcriptional regulation in the presence of several exogenous ligands. CITED2 also increased PPARγ-dependent regulation of reporter genes but had no effect on PPARβ activity. To determine whether CITED2 affects endogenous gene expression, this protein was stably overexpressed (CITED2+) or repressed by small inhibitor RNA (CITED2-) in immortalized mouse hepatocytes. Relative to the control stably transfected or CITED2- cells, CITED2+ cells had an increased rate of cell proliferation. Microarray analysis and real time PCR showed that several genes are differentially affected by PPARα ligands in CITED2+ versus CITED2- cells. Genes that were affected by PPARα ligands in a CITED2-modulatory manner include angiopoietin-like protein 4, forkhead C2, hypoxia-inducible factor-1α, and MAPK phosphatase 1. Interestingly these genes share common functions in that they are known to promote vascularization and angiogenesis in response to hypoxia. The results described here suggest that CIT-ED2 is a coactivator of PPARα and that both proteins may participate in signaling cascades of hypoxic response and angiogenesis.",
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Identification of the CREB-binding protein/p300-interacting protein CITED2 as a peroxisome proliferator-activated receptor α coregulator. / Tien, Eric S.; Davis, John W.; Vanden Heuvel, John P.

In: Journal of Biological Chemistry, Vol. 279, No. 23, 04.06.2004, p. 24053-24063.

Research output: Contribution to journalArticle

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T1 - Identification of the CREB-binding protein/p300-interacting protein CITED2 as a peroxisome proliferator-activated receptor α coregulator

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N2 - Like other nuclear receptors, the peroxisome proliferator-activated receptors (PPARs) use a wide variety of protein-protein interactions to properly regulate transcription of target genes. In an attempt to identify novel PPAR-interacting proteins, a cDNA expression library was screened with bacterially expressed PPARα. One of the genes identified as a PPARα-associated protein by interaction cloning was the CREB-binding protein/p300-interacting transactivator with ED-rich tail 2 (CITED2, also called p35srj/mrg1/msg1). This coactivator interacted directly with PPARα in the presence or absence of ligand predominantly via the ligand binding domain of the nuclear receptor. In transient transfection reporter assays, CITED2 acted as a dose-dependent coactivator of PPARα-dependent transcriptional regulation in the presence of several exogenous ligands. CITED2 also increased PPARγ-dependent regulation of reporter genes but had no effect on PPARβ activity. To determine whether CITED2 affects endogenous gene expression, this protein was stably overexpressed (CITED2+) or repressed by small inhibitor RNA (CITED2-) in immortalized mouse hepatocytes. Relative to the control stably transfected or CITED2- cells, CITED2+ cells had an increased rate of cell proliferation. Microarray analysis and real time PCR showed that several genes are differentially affected by PPARα ligands in CITED2+ versus CITED2- cells. Genes that were affected by PPARα ligands in a CITED2-modulatory manner include angiopoietin-like protein 4, forkhead C2, hypoxia-inducible factor-1α, and MAPK phosphatase 1. Interestingly these genes share common functions in that they are known to promote vascularization and angiogenesis in response to hypoxia. The results described here suggest that CIT-ED2 is a coactivator of PPARα and that both proteins may participate in signaling cascades of hypoxic response and angiogenesis.

AB - Like other nuclear receptors, the peroxisome proliferator-activated receptors (PPARs) use a wide variety of protein-protein interactions to properly regulate transcription of target genes. In an attempt to identify novel PPAR-interacting proteins, a cDNA expression library was screened with bacterially expressed PPARα. One of the genes identified as a PPARα-associated protein by interaction cloning was the CREB-binding protein/p300-interacting transactivator with ED-rich tail 2 (CITED2, also called p35srj/mrg1/msg1). This coactivator interacted directly with PPARα in the presence or absence of ligand predominantly via the ligand binding domain of the nuclear receptor. In transient transfection reporter assays, CITED2 acted as a dose-dependent coactivator of PPARα-dependent transcriptional regulation in the presence of several exogenous ligands. CITED2 also increased PPARγ-dependent regulation of reporter genes but had no effect on PPARβ activity. To determine whether CITED2 affects endogenous gene expression, this protein was stably overexpressed (CITED2+) or repressed by small inhibitor RNA (CITED2-) in immortalized mouse hepatocytes. Relative to the control stably transfected or CITED2- cells, CITED2+ cells had an increased rate of cell proliferation. Microarray analysis and real time PCR showed that several genes are differentially affected by PPARα ligands in CITED2+ versus CITED2- cells. Genes that were affected by PPARα ligands in a CITED2-modulatory manner include angiopoietin-like protein 4, forkhead C2, hypoxia-inducible factor-1α, and MAPK phosphatase 1. Interestingly these genes share common functions in that they are known to promote vascularization and angiogenesis in response to hypoxia. The results described here suggest that CIT-ED2 is a coactivator of PPARα and that both proteins may participate in signaling cascades of hypoxic response and angiogenesis.

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