LPS-induced G-CSF expression in macrophages is mediated by ERK2, but not ERK1

Shwu Fen Chang, Shih Shan Lin, Hui Ching Yang, Yuan Yi Chou, Jhen I. Gao, Shao Chun Lu

研究成果: 雜誌貢獻文章

7 引文 (Scopus)

摘要

Granulocyte colony-stimulating factor (G-CSF) selectively stimulates proliferation and differentiation of neutrophil progenitors which play important roles in host defense against infectious agents. However, persistent G-CSF production often leads to neutrophilia and excessive inflammatory reactions. There is therefore a need to understand the mechanism regulating G-CSF expression. In this study, we showed that U0126, a MEK1/2 inhibitor, decreases lipopolysaccharide (LPS)-stimulated G-CSF promoter activity, mRNA expression and protein secretion. Using short hairpin RNA knockdown, we demonstrated that ERK2, and not ERK1, involves in LPS-induced G-CSF expression, but not LPS-regulated expression of TNF-α. Reporter assays showed that ERK2 and C/EBPβ synergistically activate G-CSF promoter activity. Further chromatin immunoprecipitation (ChIP) assays revealed that U0126 inhibits LPS-induced binding of NF-κB (p50/p65) and C/EBPβ to the G-CSF promoter, but not their nuclear protein levels. Knockdown of ERK2 inhibits LPS-induced accessibility of the G-CSF promoter region to DNase I, suggesting that chromatin remodeling may occur. These findings clarify that ERK2, rather than ERK1, mediates LPS-induced G-CSF expression in macrophages by remodeling chromatin, and stimulates C/ EBPβ-dependent activation of the G-CSF promoter. This study provides a potential target for regulating G-CSF expression.

原文英語
文章編號e0129685
期刊PLoS One
10
發行號6
DOIs
出版狀態已發佈 - 六月 26 2015

指紋

granulocyte colony-stimulating factor
Macrophages
Granulocyte Colony-Stimulating Factor
lipopolysaccharides
Lipopolysaccharides
macrophages
promoter regions
Chromatin
chromatin
Chromatin Assembly and Disassembly
Assays
deoxyribonuclease I
protein secretion
Chromatin Immunoprecipitation
Deoxyribonuclease I
nuclear proteins
production economics
assays
Nuclear Proteins
small interfering RNA

ASJC Scopus subject areas

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

引用此文

Chang, S. F., Lin, S. S., Yang, H. C., Chou, Y. Y., Gao, J. I., & Lu, S. C. (2015). LPS-induced G-CSF expression in macrophages is mediated by ERK2, but not ERK1. PLoS One, 10(6), [e0129685]. https://doi.org/10.1371/journal.pone.0129685

LPS-induced G-CSF expression in macrophages is mediated by ERK2, but not ERK1. / Chang, Shwu Fen; Lin, Shih Shan; Yang, Hui Ching; Chou, Yuan Yi; Gao, Jhen I.; Lu, Shao Chun.

於: PLoS One, 卷 10, 編號 6, e0129685, 26.06.2015.

研究成果: 雜誌貢獻文章

Chang, Shwu Fen ; Lin, Shih Shan ; Yang, Hui Ching ; Chou, Yuan Yi ; Gao, Jhen I. ; Lu, Shao Chun. / LPS-induced G-CSF expression in macrophages is mediated by ERK2, but not ERK1. 於: PLoS One. 2015 ; 卷 10, 編號 6.
@article{3fd1a434b55d4b7f8010b188e568ce27,
title = "LPS-induced G-CSF expression in macrophages is mediated by ERK2, but not ERK1",
abstract = "Granulocyte colony-stimulating factor (G-CSF) selectively stimulates proliferation and differentiation of neutrophil progenitors which play important roles in host defense against infectious agents. However, persistent G-CSF production often leads to neutrophilia and excessive inflammatory reactions. There is therefore a need to understand the mechanism regulating G-CSF expression. In this study, we showed that U0126, a MEK1/2 inhibitor, decreases lipopolysaccharide (LPS)-stimulated G-CSF promoter activity, mRNA expression and protein secretion. Using short hairpin RNA knockdown, we demonstrated that ERK2, and not ERK1, involves in LPS-induced G-CSF expression, but not LPS-regulated expression of TNF-α. Reporter assays showed that ERK2 and C/EBPβ synergistically activate G-CSF promoter activity. Further chromatin immunoprecipitation (ChIP) assays revealed that U0126 inhibits LPS-induced binding of NF-κB (p50/p65) and C/EBPβ to the G-CSF promoter, but not their nuclear protein levels. Knockdown of ERK2 inhibits LPS-induced accessibility of the G-CSF promoter region to DNase I, suggesting that chromatin remodeling may occur. These findings clarify that ERK2, rather than ERK1, mediates LPS-induced G-CSF expression in macrophages by remodeling chromatin, and stimulates C/ EBPβ-dependent activation of the G-CSF promoter. This study provides a potential target for regulating G-CSF expression.",
author = "Chang, {Shwu Fen} and Lin, {Shih Shan} and Yang, {Hui Ching} and Chou, {Yuan Yi} and Gao, {Jhen I.} and Lu, {Shao Chun}",
year = "2015",
month = "6",
day = "26",
doi = "10.1371/journal.pone.0129685",
language = "English",
volume = "10",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "6",

}

TY - JOUR

T1 - LPS-induced G-CSF expression in macrophages is mediated by ERK2, but not ERK1

AU - Chang, Shwu Fen

AU - Lin, Shih Shan

AU - Yang, Hui Ching

AU - Chou, Yuan Yi

AU - Gao, Jhen I.

AU - Lu, Shao Chun

PY - 2015/6/26

Y1 - 2015/6/26

N2 - Granulocyte colony-stimulating factor (G-CSF) selectively stimulates proliferation and differentiation of neutrophil progenitors which play important roles in host defense against infectious agents. However, persistent G-CSF production often leads to neutrophilia and excessive inflammatory reactions. There is therefore a need to understand the mechanism regulating G-CSF expression. In this study, we showed that U0126, a MEK1/2 inhibitor, decreases lipopolysaccharide (LPS)-stimulated G-CSF promoter activity, mRNA expression and protein secretion. Using short hairpin RNA knockdown, we demonstrated that ERK2, and not ERK1, involves in LPS-induced G-CSF expression, but not LPS-regulated expression of TNF-α. Reporter assays showed that ERK2 and C/EBPβ synergistically activate G-CSF promoter activity. Further chromatin immunoprecipitation (ChIP) assays revealed that U0126 inhibits LPS-induced binding of NF-κB (p50/p65) and C/EBPβ to the G-CSF promoter, but not their nuclear protein levels. Knockdown of ERK2 inhibits LPS-induced accessibility of the G-CSF promoter region to DNase I, suggesting that chromatin remodeling may occur. These findings clarify that ERK2, rather than ERK1, mediates LPS-induced G-CSF expression in macrophages by remodeling chromatin, and stimulates C/ EBPβ-dependent activation of the G-CSF promoter. This study provides a potential target for regulating G-CSF expression.

AB - Granulocyte colony-stimulating factor (G-CSF) selectively stimulates proliferation and differentiation of neutrophil progenitors which play important roles in host defense against infectious agents. However, persistent G-CSF production often leads to neutrophilia and excessive inflammatory reactions. There is therefore a need to understand the mechanism regulating G-CSF expression. In this study, we showed that U0126, a MEK1/2 inhibitor, decreases lipopolysaccharide (LPS)-stimulated G-CSF promoter activity, mRNA expression and protein secretion. Using short hairpin RNA knockdown, we demonstrated that ERK2, and not ERK1, involves in LPS-induced G-CSF expression, but not LPS-regulated expression of TNF-α. Reporter assays showed that ERK2 and C/EBPβ synergistically activate G-CSF promoter activity. Further chromatin immunoprecipitation (ChIP) assays revealed that U0126 inhibits LPS-induced binding of NF-κB (p50/p65) and C/EBPβ to the G-CSF promoter, but not their nuclear protein levels. Knockdown of ERK2 inhibits LPS-induced accessibility of the G-CSF promoter region to DNase I, suggesting that chromatin remodeling may occur. These findings clarify that ERK2, rather than ERK1, mediates LPS-induced G-CSF expression in macrophages by remodeling chromatin, and stimulates C/ EBPβ-dependent activation of the G-CSF promoter. This study provides a potential target for regulating G-CSF expression.

UR - http://www.scopus.com/inward/record.url?scp=84938499450&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84938499450&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0129685

DO - 10.1371/journal.pone.0129685

M3 - Article

C2 - 26114754

AN - SCOPUS:84938499450

VL - 10

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 6

M1 - e0129685

ER -