Molecular cloning of the human platelet-derived growth factor receptor β (PDGFR-β) promoter and drug targeting of the g-quadruplex-forming region to repress PDGFR-β expression

Yong Qin, Jessica S. Fortin, Denise Tye, Mary Gleason-Guzman, Tracy A. Brooks, Laurence Hurley

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

To understand the mechanisms controlling platelet-derived growth factor receptor β (PDGFR-β) expression in malignancies, we have cloned and characterized the first functional promoter of the human PDGFR-β gene, which has been confirmed by luciferase reporter gene assays. The transcription initiation sites were mapped by primer extension. Promoter deletion experiments demonstrate that the proximal, highly GC-rich region (positions -165 to -139) of the human PDGFR-β promoter is crucial for basal promoter activity. This region is sensitive to S1 nuclease and likely to assume a non-B-form DNA secondary structure within the supercoiled plasmid. The G-rich strand in this region contains a series of runs of three or more guanines that can form multiple different G-quadruplex structures, which have been subsequently assessed by circular dichroism. A Taq polymerase stop assay has shown that three different G-quadruplex-interactive drugs can each selectively stabilize different G-quadruplex structures of the human PDGFR-β promoter. However, in transfection experiments, only telomestatin significantly reduced the human PDGFR-β basal promoter activity relative to the control. Furthermore, the PDGFR-β mRNA level in Daoy cells was significantly decreased after treatment with 1 μM telomestatin for 24 h. Therefore, we propose that ligand-mediated stabilization of specific G-quadruplex structures in the human PDGFR-β promoter can modulate its transcription.

Original languageEnglish (US)
Pages (from-to)4208-4219
Number of pages12
JournalBiochemistry
Volume49
Issue number19
DOIs
StatePublished - May 18 2010

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Platelet-Derived Growth Factor Receptors
Cloning
G-Quadruplexes
Molecular Cloning
Drug Delivery Systems
Assays
Genes
GC Rich Sequence
Taq Polymerase
Transcription Initiation Site
Guanine
Transcription
Luciferases
Plasmids
Stabilization
Experiments
Ligands
Messenger RNA
DNA
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Biochemistry

Cite this

Molecular cloning of the human platelet-derived growth factor receptor β (PDGFR-β) promoter and drug targeting of the g-quadruplex-forming region to repress PDGFR-β expression. / Qin, Yong; Fortin, Jessica S.; Tye, Denise; Gleason-Guzman, Mary; Brooks, Tracy A.; Hurley, Laurence.

In: Biochemistry, Vol. 49, No. 19, 18.05.2010, p. 4208-4219.

Research output: Contribution to journalArticle

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