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Research ArticleArticle

Regulation of the Rat Phenylethanolamine N-Methyltransferase Gene by Transcription Factors Sp1 and MAZ

Song Her, Robert Claycomb, T. C. Tai and Dona L. Wong
Molecular Pharmacology November 2003, 64 (5) 1180-1188; DOI: https://doi.org/10.1124/mol.64.5.1180
Song Her
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Robert Claycomb
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T. C. Tai
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Dona L. Wong
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Abstract

The rat phenylethanolamine N-methyltransferase (PNMT) gene promoter contains 1-base pair (bp) overlapping consensus sequences for Sp1 and MAZ transcription factors at -48 and -38 bp, respectively. Gel mobility assays using PC-12-derived RS1 cell nuclear extracts or in vitro translated proteins showed that Sp1 and MAZ specifically bind to these elements, that MAZ displaces/prevents Sp1 binding, and that Sp1 and MAZ binding is mutually exclusive, with occupancy dependent on each factor's concentration and affinity for its consensus element. In transfection assays, PNMT promoter activation by Sp1 and MAZ depends on promoter length, with -893 bp of sequence yielding greatest activation. Although MAZ has higher affinity for its binding element, it is a less effective activator. Changes in PNMT promoter activity for the constructs pGL3RP60 or pGL3RP893 using a fixed amount of MAZ expression construct and a variable amount of Sp1 expression construct or vice versa confirmed the latter. Mutation of the MAZ or Sp1 sites in pGL3RP60 attenuated but did not eliminate PNMT promoter activity, even though the proteins no longer bind to their consensus elements. Phosphatase treatment of RS1 cell nuclear extracts prevented MAZ- and Sp1-DNA binding complex formation. Although MAZ and Sp1 elevate endogenous PNMT mRNA in RS1 cells, MAZ preferentially increases intron-retaining whereas Sp1 preferentially increases intronless mRNA. Thus, expression of the PNMT gene seems to be modulated through competitive binding of phosphorylated Sp1 and MAZ to their consensus elements in the promoter. In addition, post-transcriptional regulation seems to be another important mechanism controlling PNMT expression.

  • Received April 2, 2003.
  • Accepted August 11, 2003.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 64 (5)
Molecular Pharmacology
Vol. 64, Issue 5
1 Nov 2003
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Research ArticleArticle

Regulation of the Rat Phenylethanolamine N-Methyltransferase Gene by Transcription Factors Sp1 and MAZ

Song Her, Robert Claycomb, T. C. Tai and Dona L. Wong
Molecular Pharmacology November 1, 2003, 64 (5) 1180-1188; DOI: https://doi.org/10.1124/mol.64.5.1180

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Research ArticleArticle

Regulation of the Rat Phenylethanolamine N-Methyltransferase Gene by Transcription Factors Sp1 and MAZ

Song Her, Robert Claycomb, T. C. Tai and Dona L. Wong
Molecular Pharmacology November 1, 2003, 64 (5) 1180-1188; DOI: https://doi.org/10.1124/mol.64.5.1180
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