Identification of an Enhancer That Increases miR-200b~200a~429 Gene Expression in Breast Cancer Cells

Joanne L. Attema, Andrew G. Bert, Yat Yuen Lim, Natasha Kolesnikoff, David M. Lawrence, Katherine A. Pillman, Eric Smith, Paul A. Drew, Yeesim Khew-Goodall, Frances Shannon, Gregory J. Goodall

Research output: Contribution to journalArticle

14 Citations (Scopus)
4 Downloads (Pure)

Abstract

The miR-200b similar to 200a similar to 429 gene cluster is a key regulator of EMT and cancer metastasis, however the transcription-based mechanisms controlling its expression during this process are not well understood. We have analyzed the miR-200b similar to 200a similar to 429 locus for epigenetic modifications in breast epithelial and mesenchymal cell lines using chromatin immunoprecipitation assays and DNA methylation analysis. We discovered a novel enhancer located approximately 5.1kb upstream of the miR-200b similar to 200a similar to 429 transcriptional start site. This region was associated with the active enhancer chromatin signature comprising H3K4me1, H3K27ac, RNA polymerase II and CpG dinucleotide hypomethylation. Luciferase reporter assays revealed the upstream enhancer stimulated the transcription of the miR-200b similar to 200a similar to 429 minimal promoter region approximately 27-fold in breast epithelial cells. Furthermore, we found that a region of the enhancer was transcribed, producing a short, GC-rich, mainly nuclear, non-polyadenylated RNA transcript designated miR-200b eRNA. Over-expression of miR-200b eRNA had little effect on miR-200b similar to 200a similar to 429 promoter activity and its production did not correlate with miR-200b similar to 200a similar to 429 gene expression. While additional investigations of miR-200b eRNA function will be necessary, it is possible that miR-200b eRNA may be involved in the regulation of miR-200b similar to 200a similar to 429 gene expression and silencing. Taken together, these findings reveal the presence of a novel enhancer, which contributes to miR-200b similar to 200a similar to 429 transcriptional regulation in epithelial cells
Original languageEnglish
Article numbere75517
Pages (from-to)1-15
Number of pages15
JournalPLoS One
Volume8
Issue number9
DOIs
Publication statusPublished - 2013

Fingerprint

Transcription
Gene expression
breast neoplasms
Chromatin
breasts
chromatin
Assays
epithelial cells
transcription (genetics)
Epithelial Cells
Cells
promoter regions
Breast Neoplasms
Gene Expression
gene expression
RNA Polymerase II
Breast
assays
gene silencing
DNA methylation

Cite this

Attema, J. L., Bert, A. G., Lim, Y. Y., Kolesnikoff, N., Lawrence, D. M., Pillman, K. A., ... Goodall, G. J. (2013). Identification of an Enhancer That Increases miR-200b~200a~429 Gene Expression in Breast Cancer Cells. PLoS One, 8(9), 1-15. [e75517]. https://doi.org/10.1371/journal.pone.0075517
Attema, Joanne L. ; Bert, Andrew G. ; Lim, Yat Yuen ; Kolesnikoff, Natasha ; Lawrence, David M. ; Pillman, Katherine A. ; Smith, Eric ; Drew, Paul A. ; Khew-Goodall, Yeesim ; Shannon, Frances ; Goodall, Gregory J. / Identification of an Enhancer That Increases miR-200b~200a~429 Gene Expression in Breast Cancer Cells. In: PLoS One. 2013 ; Vol. 8, No. 9. pp. 1-15.
@article{7fc478c59a2b4803b2ceec6b6c3e1fc4,
title = "Identification of an Enhancer That Increases miR-200b~200a~429 Gene Expression in Breast Cancer Cells",
abstract = "The miR-200b similar to 200a similar to 429 gene cluster is a key regulator of EMT and cancer metastasis, however the transcription-based mechanisms controlling its expression during this process are not well understood. We have analyzed the miR-200b similar to 200a similar to 429 locus for epigenetic modifications in breast epithelial and mesenchymal cell lines using chromatin immunoprecipitation assays and DNA methylation analysis. We discovered a novel enhancer located approximately 5.1kb upstream of the miR-200b similar to 200a similar to 429 transcriptional start site. This region was associated with the active enhancer chromatin signature comprising H3K4me1, H3K27ac, RNA polymerase II and CpG dinucleotide hypomethylation. Luciferase reporter assays revealed the upstream enhancer stimulated the transcription of the miR-200b similar to 200a similar to 429 minimal promoter region approximately 27-fold in breast epithelial cells. Furthermore, we found that a region of the enhancer was transcribed, producing a short, GC-rich, mainly nuclear, non-polyadenylated RNA transcript designated miR-200b eRNA. Over-expression of miR-200b eRNA had little effect on miR-200b similar to 200a similar to 429 promoter activity and its production did not correlate with miR-200b similar to 200a similar to 429 gene expression. While additional investigations of miR-200b eRNA function will be necessary, it is possible that miR-200b eRNA may be involved in the regulation of miR-200b similar to 200a similar to 429 gene expression and silencing. Taken together, these findings reveal the presence of a novel enhancer, which contributes to miR-200b similar to 200a similar to 429 transcriptional regulation in epithelial cells",
keywords = "(blank)",
author = "Attema, {Joanne L.} and Bert, {Andrew G.} and Lim, {Yat Yuen} and Natasha Kolesnikoff and Lawrence, {David M.} and Pillman, {Katherine A.} and Eric Smith and Drew, {Paul A.} and Yeesim Khew-Goodall and Frances Shannon and Goodall, {Gregory J.}",
year = "2013",
doi = "10.1371/journal.pone.0075517",
language = "English",
volume = "8",
pages = "1--15",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "9",

}

Attema, JL, Bert, AG, Lim, YY, Kolesnikoff, N, Lawrence, DM, Pillman, KA, Smith, E, Drew, PA, Khew-Goodall, Y, Shannon, F & Goodall, GJ 2013, 'Identification of an Enhancer That Increases miR-200b~200a~429 Gene Expression in Breast Cancer Cells', PLoS One, vol. 8, no. 9, e75517, pp. 1-15. https://doi.org/10.1371/journal.pone.0075517

Identification of an Enhancer That Increases miR-200b~200a~429 Gene Expression in Breast Cancer Cells. / Attema, Joanne L.; Bert, Andrew G.; Lim, Yat Yuen; Kolesnikoff, Natasha; Lawrence, David M.; Pillman, Katherine A.; Smith, Eric; Drew, Paul A.; Khew-Goodall, Yeesim; Shannon, Frances; Goodall, Gregory J.

In: PLoS One, Vol. 8, No. 9, e75517, 2013, p. 1-15.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Identification of an Enhancer That Increases miR-200b~200a~429 Gene Expression in Breast Cancer Cells

AU - Attema, Joanne L.

AU - Bert, Andrew G.

AU - Lim, Yat Yuen

AU - Kolesnikoff, Natasha

AU - Lawrence, David M.

AU - Pillman, Katherine A.

AU - Smith, Eric

AU - Drew, Paul A.

AU - Khew-Goodall, Yeesim

AU - Shannon, Frances

AU - Goodall, Gregory J.

PY - 2013

Y1 - 2013

N2 - The miR-200b similar to 200a similar to 429 gene cluster is a key regulator of EMT and cancer metastasis, however the transcription-based mechanisms controlling its expression during this process are not well understood. We have analyzed the miR-200b similar to 200a similar to 429 locus for epigenetic modifications in breast epithelial and mesenchymal cell lines using chromatin immunoprecipitation assays and DNA methylation analysis. We discovered a novel enhancer located approximately 5.1kb upstream of the miR-200b similar to 200a similar to 429 transcriptional start site. This region was associated with the active enhancer chromatin signature comprising H3K4me1, H3K27ac, RNA polymerase II and CpG dinucleotide hypomethylation. Luciferase reporter assays revealed the upstream enhancer stimulated the transcription of the miR-200b similar to 200a similar to 429 minimal promoter region approximately 27-fold in breast epithelial cells. Furthermore, we found that a region of the enhancer was transcribed, producing a short, GC-rich, mainly nuclear, non-polyadenylated RNA transcript designated miR-200b eRNA. Over-expression of miR-200b eRNA had little effect on miR-200b similar to 200a similar to 429 promoter activity and its production did not correlate with miR-200b similar to 200a similar to 429 gene expression. While additional investigations of miR-200b eRNA function will be necessary, it is possible that miR-200b eRNA may be involved in the regulation of miR-200b similar to 200a similar to 429 gene expression and silencing. Taken together, these findings reveal the presence of a novel enhancer, which contributes to miR-200b similar to 200a similar to 429 transcriptional regulation in epithelial cells

AB - The miR-200b similar to 200a similar to 429 gene cluster is a key regulator of EMT and cancer metastasis, however the transcription-based mechanisms controlling its expression during this process are not well understood. We have analyzed the miR-200b similar to 200a similar to 429 locus for epigenetic modifications in breast epithelial and mesenchymal cell lines using chromatin immunoprecipitation assays and DNA methylation analysis. We discovered a novel enhancer located approximately 5.1kb upstream of the miR-200b similar to 200a similar to 429 transcriptional start site. This region was associated with the active enhancer chromatin signature comprising H3K4me1, H3K27ac, RNA polymerase II and CpG dinucleotide hypomethylation. Luciferase reporter assays revealed the upstream enhancer stimulated the transcription of the miR-200b similar to 200a similar to 429 minimal promoter region approximately 27-fold in breast epithelial cells. Furthermore, we found that a region of the enhancer was transcribed, producing a short, GC-rich, mainly nuclear, non-polyadenylated RNA transcript designated miR-200b eRNA. Over-expression of miR-200b eRNA had little effect on miR-200b similar to 200a similar to 429 promoter activity and its production did not correlate with miR-200b similar to 200a similar to 429 gene expression. While additional investigations of miR-200b eRNA function will be necessary, it is possible that miR-200b eRNA may be involved in the regulation of miR-200b similar to 200a similar to 429 gene expression and silencing. Taken together, these findings reveal the presence of a novel enhancer, which contributes to miR-200b similar to 200a similar to 429 transcriptional regulation in epithelial cells

KW - (blank)

U2 - 10.1371/journal.pone.0075517

DO - 10.1371/journal.pone.0075517

M3 - Article

VL - 8

SP - 1

EP - 15

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 9

M1 - e75517

ER -

Attema JL, Bert AG, Lim YY, Kolesnikoff N, Lawrence DM, Pillman KA et al. Identification of an Enhancer That Increases miR-200b~200a~429 Gene Expression in Breast Cancer Cells. PLoS One. 2013;8(9):1-15. e75517. https://doi.org/10.1371/journal.pone.0075517