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Platelet-derived growth factor B-chain gene expression in mesangial cells: Effect of phorbol ester on gene transcription and mRNA stability

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Abstract

We have investigated the effect of phorbol 12-myristate 13-acetate (PMA) on platelet-derived growth factor (PDGF) B-chain gene transcription as well as on mRNA stability in cultured human mesangial cells. Addition of actinomycin to cells stimulated with PMA decreases steady state levels of PDGF-B chain mRNA analysed by solution hybridization assay. PDGF-B chain gene transcription was also assayed directly by measuring elongation of transcripts in isolated nuclei followed by hybridization of labeled RNA transcripts to a cDNA encoding for PDGF-B chain. Our data show that PMA induces PDGF-B chain gene transcription by approximately 2-fold. α-Amanitin, an RNA polymerase II inhibitor, blocked transcription by more than 70%. In addition, we determined the effect of PMA on the halflife of PDGF-B chain mRNA directly by pulse chase method. In human mesangial cells, the PDGF-B chain mRNA exhibited halflife of approximately 105 min. In the presence of PMA, the halflife of PDGF-B chain mRNA was reduced to approximately 72 min. These studies indicate that regulation of PDGF-B chain gene by PMA in human mesangial cells involves a coordinate effort at the level of transcription and mRNA stability.

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Authors and Affiliations

  1. Department of Medicine, Division of Nephrology, The University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, 78284-7882, San Antonio, TX, USA

    Basant Bhandari, Kathleen Woodruff & Hanna E. Abboud

  2. Audie L. Murphy VA Hospital, 7703 Floyd Curl Drive, 78284-7882, San Antonio, TX, USA

    Basant Bhandari, Kathleen Woodruff & Hanna E. Abboud

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  1. Basant Bhandari
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  2. Kathleen Woodruff
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  3. Hanna E. Abboud
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Bhandari, B., Woodruff, K. & Abboud, H.E. Platelet-derived growth factor B-chain gene expression in mesangial cells: Effect of phorbol ester on gene transcription and mRNA stability. Mol Cell Biochem 140, 31–36 (1994). https://doi.org/10.1007/BF00928363

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  • DOI: https://doi.org/10.1007/BF00928363

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