Transcriptional and Epigenetic Regulation of KIAA1199 Gene Expression in Human Breast Cancer

Cem Kuscu, Nikki Evensen, Deborah Kim, You Jun Hu, Stanley Zucker, Jian Cao

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Emerging evidence has demonstrated that upregulated expression of KIAA1199 in human cancer bodes for poor survival. The regulatory mechanism controlling KIAA1199 expression in cancer remains to be characterized. In the present study, we have isolated and characterized the human KIAA1199 promoter in terms of regulation of KIAA1199 gene expression. A 3.3 kb fragment of human genomic DNA containing the 5′-flanking sequence of the KIAA1199 gene possesses both suppressive and activating elements. Employing a deletion mutagenesis approach, a 1.4 kb proximal region was defined as the basic KIAA1199 promoter containing a TATA-box close to the transcription start site. A combination of 5′-primer extension study with 5′RACE DNA sequencing analysis revealed one major transcription start site that is utilized in the human KIAA1199 gene. Bioinformatics analysis suggested that the 1.4 kb KIAA1199 promoter contains putative activating regulatory elements, including activator protein-1(AP-1), Twist-1, and NF-κB sites. Sequential deletion and site-direct mutagenesis analysis demonstrated that the AP-1 and distal NF-κB sites are required for KIAA1199 gene expression. Further analyses using an electrophoretic mobility-shift assay and chromatin immunoprecipitation confirmed the requirement of these cis- and trans-acting elements in controlling KIAA1199 gene expression. Finally, we found that upregulated KIAA1199 expression in human breast cancer specimens correlated with hypomethylation of the regulatory region. Involvement of DNA methylation in regulation of KIAA1199 expression was recapitulated in human breast cancer cell lines. Taken together, our study unraveled the regulatory mechanisms controlling KIAA1199 gene expression in human cancer.

Original languageEnglish (US)
Article numbere44661
JournalPloS one
Volume7
Issue number9
DOIs
StatePublished - Sep 6 2012
Externally publishedYes

Fingerprint

gene expression regulation
Gene Expression Regulation
Epigenomics
Gene expression
epigenetics
breast neoplasms
Breast Neoplasms
Mutagenesis
Transcription Initiation Site
Transcription Factor AP-1
Genes
promoter regions
Electrophoretic mobility
TATA Box
5' Flanking Region
Nucleic Acid Regulatory Sequences
Gene Expression
DNA
mutagenesis
gene expression

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Transcriptional and Epigenetic Regulation of KIAA1199 Gene Expression in Human Breast Cancer. / Kuscu, Cem; Evensen, Nikki; Kim, Deborah; Hu, You Jun; Zucker, Stanley; Cao, Jian.

In: PloS one, Vol. 7, No. 9, e44661, 06.09.2012.

Research output: Contribution to journalArticle

Kuscu, Cem ; Evensen, Nikki ; Kim, Deborah ; Hu, You Jun ; Zucker, Stanley ; Cao, Jian. / Transcriptional and Epigenetic Regulation of KIAA1199 Gene Expression in Human Breast Cancer. In: PloS one. 2012 ; Vol. 7, No. 9.
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