MZF-1 and DbpA interact with DNase I hypersensitive sites that correlate with expression of the human MUC1 mucin gene

Toshiyuki Shiraga, John P. Winpenny, Emma J. Carter, Victoria A. McCarthy, Michael A Hollingsworth, Ann Harris

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The MUC1 mucin is a large membrane-tethered glycoprotein that shows differential expression in many adenocarcinomas, where it contributes to their invasive and metastatic properties. We previously identified DNase I hypersensitive sites at -750 and -250 bp in the human MUC1 gene promoter and showed concordance between the -250 site and MUC1 mRNA levels in vivo. Transient expression assays using promoter constructs, in which the core DHS was deleted, to drive reporter gene expression revealed in vivo evidence for their activity. DNase I footprinting using nuclear extracts from HPAF human pancreatic carcinoma cells and MCF7 breast carcinoma cells identified three protein-binding elements in these regions (-250FP1, FP2 and -750FP). Electrophoretic mobility shift assays detected several complexes between HPAF nuclear proteins and labeled FP DNA probes. Southwestern blots and UV cross-linking experiments identified myeloid zinc finger-1 (MZF-1) as a candidate transcription factor among proteins binding to the -250FP1 and FP2 sequences. Another candidate that was identified by screening an HPAF cDNA expression library with the -250FP1 probe is DNA binding protein A (DbpA). Exogenous DbpA expression in COS-7 cells was accompanied by upregulation of MUC1 promoter activity via the -250 DHS, suggesting that DbpA binding to the -250 DHS can influence human MUC1 gene expression.

Original languageEnglish (US)
Pages (from-to)41-52
Number of pages12
JournalExperimental Cell Research
Volume308
Issue number1
DOIs
StatePublished - Aug 1 2005

Fingerprint

Deoxyribonuclease I
Zinc Fingers
Staphylococcal Protein A
DNA-Binding Proteins
Mucins
Protein Binding
Southwestern Blotting
Genes
Gene Expression
COS Cells
Membrane Glycoproteins
MCF-7 Cells
DNA Probes
Electrophoretic Mobility Shift Assay
Nuclear Proteins
Gene Library
Reporter Genes
Adenocarcinoma
Transcription Factors
Up-Regulation

Keywords

  • DNase I hypersensitive sites
  • DbpA
  • HPAF
  • MUC1 gene regulation
  • MZF-1

ASJC Scopus subject areas

  • Cell Biology

Cite this

MZF-1 and DbpA interact with DNase I hypersensitive sites that correlate with expression of the human MUC1 mucin gene. / Shiraga, Toshiyuki; Winpenny, John P.; Carter, Emma J.; McCarthy, Victoria A.; Hollingsworth, Michael A; Harris, Ann.

In: Experimental Cell Research, Vol. 308, No. 1, 01.08.2005, p. 41-52.

Research output: Contribution to journalArticle

Shiraga, Toshiyuki ; Winpenny, John P. ; Carter, Emma J. ; McCarthy, Victoria A. ; Hollingsworth, Michael A ; Harris, Ann. / MZF-1 and DbpA interact with DNase I hypersensitive sites that correlate with expression of the human MUC1 mucin gene. In: Experimental Cell Research. 2005 ; Vol. 308, No. 1. pp. 41-52.
@article{4550b13fed4d4462b2ca8a920be64064,
title = "MZF-1 and DbpA interact with DNase I hypersensitive sites that correlate with expression of the human MUC1 mucin gene",
abstract = "The MUC1 mucin is a large membrane-tethered glycoprotein that shows differential expression in many adenocarcinomas, where it contributes to their invasive and metastatic properties. We previously identified DNase I hypersensitive sites at -750 and -250 bp in the human MUC1 gene promoter and showed concordance between the -250 site and MUC1 mRNA levels in vivo. Transient expression assays using promoter constructs, in which the core DHS was deleted, to drive reporter gene expression revealed in vivo evidence for their activity. DNase I footprinting using nuclear extracts from HPAF human pancreatic carcinoma cells and MCF7 breast carcinoma cells identified three protein-binding elements in these regions (-250FP1, FP2 and -750FP). Electrophoretic mobility shift assays detected several complexes between HPAF nuclear proteins and labeled FP DNA probes. Southwestern blots and UV cross-linking experiments identified myeloid zinc finger-1 (MZF-1) as a candidate transcription factor among proteins binding to the -250FP1 and FP2 sequences. Another candidate that was identified by screening an HPAF cDNA expression library with the -250FP1 probe is DNA binding protein A (DbpA). Exogenous DbpA expression in COS-7 cells was accompanied by upregulation of MUC1 promoter activity via the -250 DHS, suggesting that DbpA binding to the -250 DHS can influence human MUC1 gene expression.",
keywords = "DNase I hypersensitive sites, DbpA, HPAF, MUC1 gene regulation, MZF-1",
author = "Toshiyuki Shiraga and Winpenny, {John P.} and Carter, {Emma J.} and McCarthy, {Victoria A.} and Hollingsworth, {Michael A} and Ann Harris",
year = "2005",
month = "8",
day = "1",
doi = "10.1016/j.yexcr.2005.04.008",
language = "English (US)",
volume = "308",
pages = "41--52",
journal = "Experimental Cell Research",
issn = "0014-4827",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - MZF-1 and DbpA interact with DNase I hypersensitive sites that correlate with expression of the human MUC1 mucin gene

AU - Shiraga, Toshiyuki

AU - Winpenny, John P.

AU - Carter, Emma J.

AU - McCarthy, Victoria A.

AU - Hollingsworth, Michael A

AU - Harris, Ann

PY - 2005/8/1

Y1 - 2005/8/1

N2 - The MUC1 mucin is a large membrane-tethered glycoprotein that shows differential expression in many adenocarcinomas, where it contributes to their invasive and metastatic properties. We previously identified DNase I hypersensitive sites at -750 and -250 bp in the human MUC1 gene promoter and showed concordance between the -250 site and MUC1 mRNA levels in vivo. Transient expression assays using promoter constructs, in which the core DHS was deleted, to drive reporter gene expression revealed in vivo evidence for their activity. DNase I footprinting using nuclear extracts from HPAF human pancreatic carcinoma cells and MCF7 breast carcinoma cells identified three protein-binding elements in these regions (-250FP1, FP2 and -750FP). Electrophoretic mobility shift assays detected several complexes between HPAF nuclear proteins and labeled FP DNA probes. Southwestern blots and UV cross-linking experiments identified myeloid zinc finger-1 (MZF-1) as a candidate transcription factor among proteins binding to the -250FP1 and FP2 sequences. Another candidate that was identified by screening an HPAF cDNA expression library with the -250FP1 probe is DNA binding protein A (DbpA). Exogenous DbpA expression in COS-7 cells was accompanied by upregulation of MUC1 promoter activity via the -250 DHS, suggesting that DbpA binding to the -250 DHS can influence human MUC1 gene expression.

AB - The MUC1 mucin is a large membrane-tethered glycoprotein that shows differential expression in many adenocarcinomas, where it contributes to their invasive and metastatic properties. We previously identified DNase I hypersensitive sites at -750 and -250 bp in the human MUC1 gene promoter and showed concordance between the -250 site and MUC1 mRNA levels in vivo. Transient expression assays using promoter constructs, in which the core DHS was deleted, to drive reporter gene expression revealed in vivo evidence for their activity. DNase I footprinting using nuclear extracts from HPAF human pancreatic carcinoma cells and MCF7 breast carcinoma cells identified three protein-binding elements in these regions (-250FP1, FP2 and -750FP). Electrophoretic mobility shift assays detected several complexes between HPAF nuclear proteins and labeled FP DNA probes. Southwestern blots and UV cross-linking experiments identified myeloid zinc finger-1 (MZF-1) as a candidate transcription factor among proteins binding to the -250FP1 and FP2 sequences. Another candidate that was identified by screening an HPAF cDNA expression library with the -250FP1 probe is DNA binding protein A (DbpA). Exogenous DbpA expression in COS-7 cells was accompanied by upregulation of MUC1 promoter activity via the -250 DHS, suggesting that DbpA binding to the -250 DHS can influence human MUC1 gene expression.

KW - DNase I hypersensitive sites

KW - DbpA

KW - HPAF

KW - MUC1 gene regulation

KW - MZF-1

UR - http://www.scopus.com/inward/record.url?scp=22144449896&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=22144449896&partnerID=8YFLogxK

U2 - 10.1016/j.yexcr.2005.04.008

DO - 10.1016/j.yexcr.2005.04.008

M3 - Article

C2 - 15893750

AN - SCOPUS:22144449896

VL - 308

SP - 41

EP - 52

JO - Experimental Cell Research

JF - Experimental Cell Research

SN - 0014-4827

IS - 1

ER -