Overlapping DNA recognition motifs between Sp1 and a novel trans-acting factor within the wt1 tumour suppressor gene promoter

Maria Teresa Discenza, Mohammed Dehbi, Jerry Pelletier

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The Wilms' tumor suppressor gene, wt1, encodes a zinc finger transcription factor which has been shown to regulate the expression of several genes involved in cellular proliferation and differentiation. Expression of wt1 is devetopmentally regulated and restricted to a small set of tissues which include the fetal urogenital system, mesothelium and spleen. A highly conserved motif within the wt1 promoter, located between nucleotides -34 and -71 relative to the first transcription start site in the murine promoter, harbors consensus binding sites for Sp1 and members of the paired-box transcription factor family. Pax-2 and Pax-8 are known to enhance expression of wt1 through this conserved regulatory element. In this report, we demonstrate that Sp1 is able to bind to two sites within the 38 bp conserved region (CR). By electrophoretic mobility shift assays (EMSAs), we have identified a novel binding activity, referred to as complex D, which recognizes sequences overlapping one of the Sp1 sites in the CR. EMSA competition experiments indicate that binding of complex D and Sp1 to the CR is mutually exclusive and Sp1 is able to displace complex D binding. In situ UV crosslinking and molecular mass determinations indicate that complex D is a complex of ≃130 kDa, consisting of at least two proteins of ≃62 and ≃70 kDa. Transient transfections suggest that complex D may function as an activator.

Original languageEnglish
Pages (from-to)4314-4322
Number of pages9
JournalNucleic Acids Research
Volume25
Issue number21
DOIs
Publication statusPublished - 1 Nov 1997
Externally publishedYes

Fingerprint

Nucleotide Motifs
Trans-Activators
Electrophoretic Mobility Shift Assay
Tumor Suppressor Genes
Paired Box Transcription Factors
Wilms' Tumor Genes
Activator Appliances
Urogenital System
Transcription Initiation Site
Zinc Fingers
Transfection
Transcription Factors
Spleen
Epithelium
Nucleotides
Binding Sites
Cell Proliferation
Gene Expression
Proteins

ASJC Scopus subject areas

  • Genetics

Cite this

Overlapping DNA recognition motifs between Sp1 and a novel trans-acting factor within the wt1 tumour suppressor gene promoter. / Discenza, Maria Teresa; Dehbi, Mohammed; Pelletier, Jerry.

In: Nucleic Acids Research, Vol. 25, No. 21, 01.11.1997, p. 4314-4322.

Research output: Contribution to journalArticle

@article{d94f34b6988f4279ac004a538839c76b,
title = "Overlapping DNA recognition motifs between Sp1 and a novel trans-acting factor within the wt1 tumour suppressor gene promoter",
abstract = "The Wilms' tumor suppressor gene, wt1, encodes a zinc finger transcription factor which has been shown to regulate the expression of several genes involved in cellular proliferation and differentiation. Expression of wt1 is devetopmentally regulated and restricted to a small set of tissues which include the fetal urogenital system, mesothelium and spleen. A highly conserved motif within the wt1 promoter, located between nucleotides -34 and -71 relative to the first transcription start site in the murine promoter, harbors consensus binding sites for Sp1 and members of the paired-box transcription factor family. Pax-2 and Pax-8 are known to enhance expression of wt1 through this conserved regulatory element. In this report, we demonstrate that Sp1 is able to bind to two sites within the 38 bp conserved region (CR). By electrophoretic mobility shift assays (EMSAs), we have identified a novel binding activity, referred to as complex D, which recognizes sequences overlapping one of the Sp1 sites in the CR. EMSA competition experiments indicate that binding of complex D and Sp1 to the CR is mutually exclusive and Sp1 is able to displace complex D binding. In situ UV crosslinking and molecular mass determinations indicate that complex D is a complex of ≃130 kDa, consisting of at least two proteins of ≃62 and ≃70 kDa. Transient transfections suggest that complex D may function as an activator.",
author = "Discenza, {Maria Teresa} and Mohammed Dehbi and Jerry Pelletier",
year = "1997",
month = "11",
day = "1",
doi = "10.1093/nar/25.21.4314",
language = "English",
volume = "25",
pages = "4314--4322",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "21",

}

TY - JOUR

T1 - Overlapping DNA recognition motifs between Sp1 and a novel trans-acting factor within the wt1 tumour suppressor gene promoter

AU - Discenza, Maria Teresa

AU - Dehbi, Mohammed

AU - Pelletier, Jerry

PY - 1997/11/1

Y1 - 1997/11/1

N2 - The Wilms' tumor suppressor gene, wt1, encodes a zinc finger transcription factor which has been shown to regulate the expression of several genes involved in cellular proliferation and differentiation. Expression of wt1 is devetopmentally regulated and restricted to a small set of tissues which include the fetal urogenital system, mesothelium and spleen. A highly conserved motif within the wt1 promoter, located between nucleotides -34 and -71 relative to the first transcription start site in the murine promoter, harbors consensus binding sites for Sp1 and members of the paired-box transcription factor family. Pax-2 and Pax-8 are known to enhance expression of wt1 through this conserved regulatory element. In this report, we demonstrate that Sp1 is able to bind to two sites within the 38 bp conserved region (CR). By electrophoretic mobility shift assays (EMSAs), we have identified a novel binding activity, referred to as complex D, which recognizes sequences overlapping one of the Sp1 sites in the CR. EMSA competition experiments indicate that binding of complex D and Sp1 to the CR is mutually exclusive and Sp1 is able to displace complex D binding. In situ UV crosslinking and molecular mass determinations indicate that complex D is a complex of ≃130 kDa, consisting of at least two proteins of ≃62 and ≃70 kDa. Transient transfections suggest that complex D may function as an activator.

AB - The Wilms' tumor suppressor gene, wt1, encodes a zinc finger transcription factor which has been shown to regulate the expression of several genes involved in cellular proliferation and differentiation. Expression of wt1 is devetopmentally regulated and restricted to a small set of tissues which include the fetal urogenital system, mesothelium and spleen. A highly conserved motif within the wt1 promoter, located between nucleotides -34 and -71 relative to the first transcription start site in the murine promoter, harbors consensus binding sites for Sp1 and members of the paired-box transcription factor family. Pax-2 and Pax-8 are known to enhance expression of wt1 through this conserved regulatory element. In this report, we demonstrate that Sp1 is able to bind to two sites within the 38 bp conserved region (CR). By electrophoretic mobility shift assays (EMSAs), we have identified a novel binding activity, referred to as complex D, which recognizes sequences overlapping one of the Sp1 sites in the CR. EMSA competition experiments indicate that binding of complex D and Sp1 to the CR is mutually exclusive and Sp1 is able to displace complex D binding. In situ UV crosslinking and molecular mass determinations indicate that complex D is a complex of ≃130 kDa, consisting of at least two proteins of ≃62 and ≃70 kDa. Transient transfections suggest that complex D may function as an activator.

UR - http://www.scopus.com/inward/record.url?scp=0030656245&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030656245&partnerID=8YFLogxK

U2 - 10.1093/nar/25.21.4314

DO - 10.1093/nar/25.21.4314

M3 - Article

VL - 25

SP - 4314

EP - 4322

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 21

ER -