title: Short nucleotide polymorphic insertions in the MCL-1 promoter affect gene expression.
abstract: We have recently reported novel short nucleotide (six and eighteen) polymorphic insertions, in the MCL-1 promoter and their association with higher mRNA and protein levels. The aim of the present study was to test the hypothesis that these insertions directly affect MCL-1 gene expression. Haematopoietic and epithelial human cell lines were transfected with +0, +6, or +18 MCL-1 promoter fragments positioned upstream of the Firefly luciferase reporter gene. The cells were stimulated with phorbol 12-myristate 13-acetate (PMA) and granulocyte macrophage colony-stimulating factor (GM-CSF). Compared to +0, both polymorphic insertions (+6 and +18) were associated with increased promoter activity. Although chromatin immunoprecipitation assay showed that there are Sp1/Sp3 binding sites in the MCL-1 promoter, electrophoretic mobility shift assay showed that it is unlikely that these sites are in the region harboring these insertions. These results provide further evidence for the biological effect of MCL-1 promoter polymorphisms on gene expression.
authors.affiliation: Department of Pathology and Laboratory Medicine, Royal University Hospital, College of Medicine, University of Saskatchewan, Saskatoon, Canada S7N 0W8. anurag.saxena@usask.ca <anurag.saxena@usask.ca>
authors.forename: Anurag
authors.initials: A
authors.lastname: Saxena
authors.forename: Oksana V
authors.initials: OV
authors.lastname: Moshynska
authors.forename: Igor D
authors.initials: ID
authors.lastname: Moshynskyy
authors.forename: Evan D
authors.initials: ED
authors.lastname: Neuls
authors.forename: Tania
authors.initials: T
authors.lastname: Qureshi
authors.forename: Mark
authors.initials: M
authors.lastname: Bosch
authors.forename: Michael
authors.initials: M
authors.lastname: Voralia
authors.forename: Keith
authors.initials: K
authors.lastname: Bonham
mesh_terms: D001483:Base Sequence; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D024202:Electrophoretic Mobility Shift Assay; D005786:Gene Expression Regulation; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D020014:K562 Cells; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D009363:Neoplasm Proteins / Q000235:genetics* / Q000378:metabolism; D009841:Oligonucleotides / Q000235:genetics* / Q000378:metabolism; D011110:Polymorphism, Genetic; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding / Q000187:drug effects; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016329:Sp1 Transcription Factor / Q000378:metabolism; D051705:Sp3 Transcription Factor / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection
chemical_list: D064549:Myeloid Cell Leukemia Sequence 1 Protein; D009363:Neoplasm Proteins; D009841:Oligonucleotides; D019253:Proto-Oncogene Proteins c-bcl-2; D011993:Recombinant Fusion Proteins; C497121:SP3 protein, human; D016329:Sp1 Transcription Factor; D051705:Sp3 Transcription Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D008156:Luciferases; D013755:Tetradecanoylphorbol Acetate