Short nucleotide polymorphic insertions in the MCL-1 promoter affect gene expression

Anurag Saxena; Oksana V Moshynska; Igor D Moshynskyy; Evan D Neuls; Tania Qureshi; Mark Bosch; Michael Voralia; Keith Bonham

doi:10.1016/j.canlet.2006.11.007

Short nucleotide polymorphic insertions in the MCL-1 promoter affect gene expression

Cancer Lett. 2007 Jun 18;251(1):114-31. doi: 10.1016/j.canlet.2006.11.007. Epub 2007 Jan 2.

Authors

Anurag Saxena¹, Oksana V Moshynska, Igor D Moshynskyy, Evan D Neuls, Tania Qureshi, Mark Bosch, Michael Voralia, Keith Bonham

Affiliation

¹ Department of Pathology and Laboratory Medicine, Royal University Hospital, College of Medicine, University of Saskatchewan, Saskatoon, Canada S7N 0W8. [email protected] <[email protected]>

PMID: 17198743
DOI: 10.1016/j.canlet.2006.11.007

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Base Sequence
Blotting, Western
Cell Line, Tumor
Cells, Cultured
Electrophoretic Mobility Shift Assay
Gene Expression Regulation*
Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
HeLa Cells
Humans
K562 Cells
Luciferases / genetics
Luciferases / metabolism
Molecular Sequence Data
Mutagenesis, Insertional*
Myeloid Cell Leukemia Sequence 1 Protein
Neoplasm Proteins / genetics*
Neoplasm Proteins / metabolism
Oligonucleotides / genetics*
Oligonucleotides / metabolism
Polymorphism, Genetic
Promoter Regions, Genetic / genetics*
Protein Binding / drug effects
Proto-Oncogene Proteins c-bcl-2 / genetics*
Proto-Oncogene Proteins c-bcl-2 / metabolism
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / metabolism
Sp1 Transcription Factor / metabolism
Sp3 Transcription Factor / metabolism
Tetradecanoylphorbol Acetate / pharmacology
Transfection

Substances

Myeloid Cell Leukemia Sequence 1 Protein
Neoplasm Proteins
Oligonucleotides
Proto-Oncogene Proteins c-bcl-2
Recombinant Fusion Proteins
SP3 protein, human
Sp1 Transcription Factor
Sp3 Transcription Factor
Granulocyte-Macrophage Colony-Stimulating Factor
Luciferases
Tetradecanoylphorbol Acetate