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Mechanism of increased PLD1 gene expression during early adipocyte differentiation process of mouse cell line 3T3-L1.
J Cell Biochem. 2010 Feb 01; 109(2):375-82.JC

Abstract

A mouse cell line 3T3-L1 is differentiated into adipocytes when treated with an inducer cocktail (IDX) (insulin, dexametahsone, and a cAMP phosphodiesterase inhibitor of isobutyl-methylxanthine (IBMX)). Here, we report that PLD1, but not PLD2, mRNA and protein increased during the early differentiation process. Our analysis shows that IDX resulted in a sequential induction of C/EBPbeta, PLD1, and C/EBPalpha which is a key transcription factor of late adipocyte differentiation. Among the three inducers, IBMX + any other inducer induced mild adipocyte differentiation, whereas insulin + dexamethasone did not. IBMX increased PLD1 but not PLD2 mRNA. Forskolin, an adenylate cyclase activator, and dbcAMP also increased PLD1 mRNA, suggesting the cellular cAMP as the inducer of both adipocyte differentiation and PLD1 transcription. We focused on the regulatory mechanism of PLD1 transcription during this differentiation process. IDX or a combination of inducers including IBMX increased PLD1 promoter activity, which is consistent with mRNA analysis. Promoter analysis identified two adjacent C/EBP motifs located between -338 and -231 bp from the first exon as the IBMX responsive elements. Furthermore, overexpression of C/EBPbeta, but not C/EBPalpha, increased PLD1 mRNA and PLD1 5' promoter activity. EMSA and chromatin immunoprecipitation assay confirmed the direct binding of C/EBPbeta, but not C/EBPalpha, to these C/EBP motifs of PLD1 5' promoter. Our results show that PLD1 is a target gene of C/EBPbeta through the increased cellular cAMP during early adipocyte differentiation of 3T3-L1 cells.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Gao S
Department of Medical Technology, Nagoya University Graduate School of Health Sciences, Nagoya, Japan.
Ito H
No affiliation info available
Murakami M
No affiliation info available
Yoshida K
No affiliation info available
Tagawa Y
No affiliation info available
Hagiwara K
No affiliation info available
Takagi A
No affiliation info available
Kojima T
No affiliation info available
Suzuki M
No affiliation info available
Banno Y
No affiliation info available
Ohguchi K
No affiliation info available
Nozawa Y
No affiliation info available
Murate T
No affiliation info available

MeSH

1-Methyl-3-isobutylxanthine3T3-L1 CellsAdipocytesAdipogenesisAnimalsCCAAT-Enhancer-Binding Protein-alphaCCAAT-Enhancer-Binding Protein-betaCyclic AMPDexamethasoneGene Expression RegulationInsulinMicePhosphodiesterase InhibitorsPhospholipase DResponse Elements

Pub Type(s)

Journal Article

Language

eng

PubMed ID

19950202

Citation

Gao, Siqiang, et al. "Mechanism of Increased PLD1 Gene Expression During Early Adipocyte Differentiation Process of Mouse Cell Line 3T3-L1." Journal of Cellular Biochemistry, vol. 109, no. 2, 2010, pp. 375-82.
Gao S, Ito H, Murakami M, et al. Mechanism of increased PLD1 gene expression during early adipocyte differentiation process of mouse cell line 3T3-L1. J Cell Biochem. 2010;109(2):375-82.
Gao, S., Ito, H., Murakami, M., Yoshida, K., Tagawa, Y., Hagiwara, K., Takagi, A., Kojima, T., Suzuki, M., Banno, Y., Ohguchi, K., Nozawa, Y., & Murate, T. (2010). Mechanism of increased PLD1 gene expression during early adipocyte differentiation process of mouse cell line 3T3-L1. Journal of Cellular Biochemistry, 109(2), 375-82. https://doi.org/10.1002/jcb.22414
Gao S, et al. Mechanism of Increased PLD1 Gene Expression During Early Adipocyte Differentiation Process of Mouse Cell Line 3T3-L1. J Cell Biochem. 2010 Feb 1;109(2):375-82. PubMed PMID: 19950202.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Mechanism of increased PLD1 gene expression during early adipocyte differentiation process of mouse cell line 3T3-L1. AU - Gao,Siqiang, AU - Ito,Hiromi, AU - Murakami,Masashi, AU - Yoshida,Kayo, AU - Tagawa,Yoko, AU - Hagiwara,Kazumi, AU - Takagi,Akira, AU - Kojima,Tetsuhito, AU - Suzuki,Motoshi, AU - Banno,Yoshiko, AU - Ohguchi,Kenji, AU - Nozawa,Yoshinori, AU - Murate,Takashi, PY - 2009/12/2/entrez PY - 2009/12/2/pubmed PY - 2010/8/6/medline SP - 375 EP - 82 JF - Journal of cellular biochemistry JO - J Cell Biochem VL - 109 IS - 2 N2 - A mouse cell line 3T3-L1 is differentiated into adipocytes when treated with an inducer cocktail (IDX) (insulin, dexametahsone, and a cAMP phosphodiesterase inhibitor of isobutyl-methylxanthine (IBMX)). Here, we report that PLD1, but not PLD2, mRNA and protein increased during the early differentiation process. Our analysis shows that IDX resulted in a sequential induction of C/EBPbeta, PLD1, and C/EBPalpha which is a key transcription factor of late adipocyte differentiation. Among the three inducers, IBMX + any other inducer induced mild adipocyte differentiation, whereas insulin + dexamethasone did not. IBMX increased PLD1 but not PLD2 mRNA. Forskolin, an adenylate cyclase activator, and dbcAMP also increased PLD1 mRNA, suggesting the cellular cAMP as the inducer of both adipocyte differentiation and PLD1 transcription. We focused on the regulatory mechanism of PLD1 transcription during this differentiation process. IDX or a combination of inducers including IBMX increased PLD1 promoter activity, which is consistent with mRNA analysis. Promoter analysis identified two adjacent C/EBP motifs located between -338 and -231 bp from the first exon as the IBMX responsive elements. Furthermore, overexpression of C/EBPbeta, but not C/EBPalpha, increased PLD1 mRNA and PLD1 5' promoter activity. EMSA and chromatin immunoprecipitation assay confirmed the direct binding of C/EBPbeta, but not C/EBPalpha, to these C/EBP motifs of PLD1 5' promoter. Our results show that PLD1 is a target gene of C/EBPbeta through the increased cellular cAMP during early adipocyte differentiation of 3T3-L1 cells. SN - 1097-4644 UR - https://www.unboundmedicine.com/medline/citation/19950202/Mechanism_of_increased_PLD1_gene_expression_during_early_adipocyte_differentiation_process_of_mouse_cell_line_3T3_L1_ L2 - https://doi.org/10.1002/jcb.22414 DB - PRIME DP - Unbound Medicine ER -