Allele-specific regulation of matrix metalloproteinase-3 gene by transcription factor NFkappaB

Veronika Souslova; Paul A Townsend; Jelena Mann; Chris M van der Loos; Anna Motterle; Fulvio D'Acquisto; Derek A Mann; Shu Ye

doi:10.1371/journal.pone.0009902

Allele-specific regulation of matrix metalloproteinase-3 gene by transcription factor NFkappaB

Veronika Souslova, Paul A Townsend, Jelena Mann, Chris M van der Loos, Anna Motterle, Fulvio D'Acquisto, Derek A Mann, Shu Ye

Research output: Contribution to journal › Article › peer-review

Abstract

BACKGROUND: Matrix metalloproteinase-3 (MMP3) is implicated in the pathogenesis and progression of atherosclerotic lesions. Previous studies suggested that MMP3 expression is influenced by a polymorphism (known as the 5A/6A polymorphism) in the promoter of the MMP3 gene and that this polymorphism is located within a cis-element that interacts with the transcription factor NFkappaB. In the present study, we sought to investigate whether MMP3 and NFkappaB were co-localized in atherosclerotic lesions and whether NFkappaB had differential effects on the two alleles of the MMP3 5A/6A polymorphism.

METHODOLOGY/PRINCIPAL FINDINGS: Immunohistochemical examination showed that MMP3 and both the NFkappaB p50 and p65 subunits were expressed abundantly in macrophages in atherosclerotic lesions and that MMP3 expression was co-localized with p50 and p65. Chromatin immunoprecipitation experiments showed interaction of p50 and p65 with the MMP3 promoter in macrophages, with greater binding to the 5A allele than to the 6A allele. Reporter gene assays in transiently transfected macrophages showed that the 5A allele had greater transcriptional activity than the 6A allele, and that this allele-specific effect was augmented when the cells were treated with the NFkappaB activator lipopolysaccharides or co-transfected with p50 and/or p65 expressing plasmids, but was reduced when the cells were treated with the NFkappaB inhibitor 6-Amino-4-(4-phenoxyphenylethylamino)-quinazoline or transfected with a dominant negative mutant of IkB kinase-beta.

CONCLUSION: These results corroborate an effect of the 5A/6A polymorphism on MMP3 transcription and indicate that NFkappaB has differential effects on the 5A and 6A alleles.

Original language	English
Pages (from-to)	e9902
Journal	PLoS One
Volume	5
Issue number	3
DOIs	https://doi.org/10.1371/journal.pone.0009902
Publication status	Published - 25 Mar 2010

Keywords

Alleles
Animals
Atherosclerosis
Gene Expression Regulation, Enzymologic
Genes, Dominant
Humans
Immunohistochemistry
Lipopolysaccharides
Matrix Metalloproteinase 3
Mice
Mutation
NF-kappa B
Polymorphism, Genetic
Promoter Regions, Genetic
Quinazolines
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

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10.1371/journal.pone.0009902

Cite this

@article{c48c962573dc4866a8cd299478ebd961,

title = "Allele-specific regulation of matrix metalloproteinase-3 gene by transcription factor NFkappaB",

abstract = "BACKGROUND: Matrix metalloproteinase-3 (MMP3) is implicated in the pathogenesis and progression of atherosclerotic lesions. Previous studies suggested that MMP3 expression is influenced by a polymorphism (known as the 5A/6A polymorphism) in the promoter of the MMP3 gene and that this polymorphism is located within a cis-element that interacts with the transcription factor NFkappaB. In the present study, we sought to investigate whether MMP3 and NFkappaB were co-localized in atherosclerotic lesions and whether NFkappaB had differential effects on the two alleles of the MMP3 5A/6A polymorphism.METHODOLOGY/PRINCIPAL FINDINGS: Immunohistochemical examination showed that MMP3 and both the NFkappaB p50 and p65 subunits were expressed abundantly in macrophages in atherosclerotic lesions and that MMP3 expression was co-localized with p50 and p65. Chromatin immunoprecipitation experiments showed interaction of p50 and p65 with the MMP3 promoter in macrophages, with greater binding to the 5A allele than to the 6A allele. Reporter gene assays in transiently transfected macrophages showed that the 5A allele had greater transcriptional activity than the 6A allele, and that this allele-specific effect was augmented when the cells were treated with the NFkappaB activator lipopolysaccharides or co-transfected with p50 and/or p65 expressing plasmids, but was reduced when the cells were treated with the NFkappaB inhibitor 6-Amino-4-(4-phenoxyphenylethylamino)-quinazoline or transfected with a dominant negative mutant of IkB kinase-beta.CONCLUSION: These results corroborate an effect of the 5A/6A polymorphism on MMP3 transcription and indicate that NFkappaB has differential effects on the 5A and 6A alleles.",

keywords = "Alleles, Animals, Atherosclerosis, Gene Expression Regulation, Enzymologic, Genes, Dominant, Humans, Immunohistochemistry, Lipopolysaccharides, Matrix Metalloproteinase 3, Mice, Mutation, NF-kappa B, Polymorphism, Genetic, Promoter Regions, Genetic, Quinazolines, Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't",

author = "Veronika Souslova and Townsend, {Paul A} and Jelena Mann and {van der Loos}, {Chris M} and Anna Motterle and Fulvio D'Acquisto and Mann, {Derek A} and Shu Ye",

year = "2010",

month = mar,

day = "25",

doi = "10.1371/journal.pone.0009902",

language = "English",

volume = "5",

pages = "e9902",

journal = "PLoS One",

issn = "1932-6203",

publisher = "Public Library of Science",

number = "3",

}

TY - JOUR

T1 - Allele-specific regulation of matrix metalloproteinase-3 gene by transcription factor NFkappaB

AU - Souslova, Veronika

AU - Townsend, Paul A

AU - Mann, Jelena

AU - van der Loos, Chris M

AU - Motterle, Anna

AU - D'Acquisto, Fulvio

AU - Mann, Derek A

AU - Ye, Shu

PY - 2010/3/25

Y1 - 2010/3/25

N2 - BACKGROUND: Matrix metalloproteinase-3 (MMP3) is implicated in the pathogenesis and progression of atherosclerotic lesions. Previous studies suggested that MMP3 expression is influenced by a polymorphism (known as the 5A/6A polymorphism) in the promoter of the MMP3 gene and that this polymorphism is located within a cis-element that interacts with the transcription factor NFkappaB. In the present study, we sought to investigate whether MMP3 and NFkappaB were co-localized in atherosclerotic lesions and whether NFkappaB had differential effects on the two alleles of the MMP3 5A/6A polymorphism.METHODOLOGY/PRINCIPAL FINDINGS: Immunohistochemical examination showed that MMP3 and both the NFkappaB p50 and p65 subunits were expressed abundantly in macrophages in atherosclerotic lesions and that MMP3 expression was co-localized with p50 and p65. Chromatin immunoprecipitation experiments showed interaction of p50 and p65 with the MMP3 promoter in macrophages, with greater binding to the 5A allele than to the 6A allele. Reporter gene assays in transiently transfected macrophages showed that the 5A allele had greater transcriptional activity than the 6A allele, and that this allele-specific effect was augmented when the cells were treated with the NFkappaB activator lipopolysaccharides or co-transfected with p50 and/or p65 expressing plasmids, but was reduced when the cells were treated with the NFkappaB inhibitor 6-Amino-4-(4-phenoxyphenylethylamino)-quinazoline or transfected with a dominant negative mutant of IkB kinase-beta.CONCLUSION: These results corroborate an effect of the 5A/6A polymorphism on MMP3 transcription and indicate that NFkappaB has differential effects on the 5A and 6A alleles.

AB - BACKGROUND: Matrix metalloproteinase-3 (MMP3) is implicated in the pathogenesis and progression of atherosclerotic lesions. Previous studies suggested that MMP3 expression is influenced by a polymorphism (known as the 5A/6A polymorphism) in the promoter of the MMP3 gene and that this polymorphism is located within a cis-element that interacts with the transcription factor NFkappaB. In the present study, we sought to investigate whether MMP3 and NFkappaB were co-localized in atherosclerotic lesions and whether NFkappaB had differential effects on the two alleles of the MMP3 5A/6A polymorphism.METHODOLOGY/PRINCIPAL FINDINGS: Immunohistochemical examination showed that MMP3 and both the NFkappaB p50 and p65 subunits were expressed abundantly in macrophages in atherosclerotic lesions and that MMP3 expression was co-localized with p50 and p65. Chromatin immunoprecipitation experiments showed interaction of p50 and p65 with the MMP3 promoter in macrophages, with greater binding to the 5A allele than to the 6A allele. Reporter gene assays in transiently transfected macrophages showed that the 5A allele had greater transcriptional activity than the 6A allele, and that this allele-specific effect was augmented when the cells were treated with the NFkappaB activator lipopolysaccharides or co-transfected with p50 and/or p65 expressing plasmids, but was reduced when the cells were treated with the NFkappaB inhibitor 6-Amino-4-(4-phenoxyphenylethylamino)-quinazoline or transfected with a dominant negative mutant of IkB kinase-beta.CONCLUSION: These results corroborate an effect of the 5A/6A polymorphism on MMP3 transcription and indicate that NFkappaB has differential effects on the 5A and 6A alleles.

KW - Alleles

KW - Animals

KW - Atherosclerosis

KW - Gene Expression Regulation, Enzymologic

KW - Genes, Dominant

KW - Humans

KW - Immunohistochemistry

KW - Lipopolysaccharides

KW - Matrix Metalloproteinase 3

KW - Mice

KW - Mutation

KW - NF-kappa B

KW - Polymorphism, Genetic

KW - Promoter Regions, Genetic

KW - Quinazolines

KW - Journal Article

KW - Research Support, N.I.H., Extramural

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

U2 - 10.1371/journal.pone.0009902

DO - 10.1371/journal.pone.0009902

M3 - Article

C2 - 20360864

SN - 1932-6203

VL - 5

SP - e9902

JO - PLoS One

JF - PLoS One

IS - 3

ER -