Immunoproteasome Deficiency Modifies the Alternative Pathway of NFκB Signaling

Marcela Maldonado, Rebecca J. Kapphahn, Marcia R. Terluk, Neal D. Heuss, Ching Yuan, Dale S. Gregerson, Deborah A. Ferrington

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Immunoproteasome is a protease abundant in immune cells and also present, albeit at lower concentrations, in cells outside the immune system. Recent evidence supports a novel role for the immunoproteasome in the cellular stress response potentially through regulation of NFκB signaling, which is the primary response to multiple stressors. The current study tests whether the Classical or Alternative Pathways are regulated by immunoproteasome following chronic TNFα exposure in cultured retinal pigment epithelial cells isolated from wild-type mice and mice deficient in one (LMP2, L2) or two (LMP7 and MECL-1, L7M1) immunoproteasome subunits. Assays were performed to assess the expression of NFκB responsive genes, the content and activity of NFκB transcription factors (p65, p50, p52, cRel, RelB), and expression and content of regulatory proteins (IκBα, A20, RPS3). Major findings include distinct differences in expression of NFκB responsive genes in both KO cells. The mechanism responsible for the altered gene expression could not be established for L7M1 since no major differences in NFκB transcription factor content or activation were observed. However, L2 cells exhibited substantially higher content and diminished activation of NFκB transcription factors associated with the Alternative Pathway and delayed termination of the Classical Pathway. These results provide strong experimental evidence supporting a role for immunoproteasome in modulating NFκB signaling.

Original languageEnglish (US)
Article numbere56187
JournalPloS one
Volume8
Issue number2
DOIs
StatePublished - Feb 14 2013

Fingerprint

Transcription Factor RelA
Transcription Factors
Genes
Chemical activation
Retinal Pigments
transcription factors
Immune system
Gene expression
Assays
Peptide Hydrolases
cells
regulatory proteins
mice
Transcriptional Activation
immune system
Immune System
stress response
Proteins
epithelial cells
proteinases

Cite this

Immunoproteasome Deficiency Modifies the Alternative Pathway of NFκB Signaling. / Maldonado, Marcela; Kapphahn, Rebecca J.; Terluk, Marcia R.; Heuss, Neal D.; Yuan, Ching; Gregerson, Dale S.; Ferrington, Deborah A.

In: PloS one, Vol. 8, No. 2, e56187, 14.02.2013.

Research output: Contribution to journalArticle

@article{25687e0b5847448e9c0f0fdce86be7ea,
title = "Immunoproteasome Deficiency Modifies the Alternative Pathway of NFκB Signaling",
abstract = "Immunoproteasome is a protease abundant in immune cells and also present, albeit at lower concentrations, in cells outside the immune system. Recent evidence supports a novel role for the immunoproteasome in the cellular stress response potentially through regulation of NFκB signaling, which is the primary response to multiple stressors. The current study tests whether the Classical or Alternative Pathways are regulated by immunoproteasome following chronic TNFα exposure in cultured retinal pigment epithelial cells isolated from wild-type mice and mice deficient in one (LMP2, L2) or two (LMP7 and MECL-1, L7M1) immunoproteasome subunits. Assays were performed to assess the expression of NFκB responsive genes, the content and activity of NFκB transcription factors (p65, p50, p52, cRel, RelB), and expression and content of regulatory proteins (IκBα, A20, RPS3). Major findings include distinct differences in expression of NFκB responsive genes in both KO cells. The mechanism responsible for the altered gene expression could not be established for L7M1 since no major differences in NFκB transcription factor content or activation were observed. However, L2 cells exhibited substantially higher content and diminished activation of NFκB transcription factors associated with the Alternative Pathway and delayed termination of the Classical Pathway. These results provide strong experimental evidence supporting a role for immunoproteasome in modulating NFκB signaling.",
author = "Marcela Maldonado and Kapphahn, {Rebecca J.} and Terluk, {Marcia R.} and Heuss, {Neal D.} and Ching Yuan and Gregerson, {Dale S.} and Ferrington, {Deborah A.}",
year = "2013",
month = "2",
day = "14",
doi = "10.1371/journal.pone.0056187",
language = "English (US)",
volume = "8",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "2",

}

TY - JOUR

T1 - Immunoproteasome Deficiency Modifies the Alternative Pathway of NFκB Signaling

AU - Maldonado, Marcela

AU - Kapphahn, Rebecca J.

AU - Terluk, Marcia R.

AU - Heuss, Neal D.

AU - Yuan, Ching

AU - Gregerson, Dale S.

AU - Ferrington, Deborah A.

PY - 2013/2/14

Y1 - 2013/2/14

N2 - Immunoproteasome is a protease abundant in immune cells and also present, albeit at lower concentrations, in cells outside the immune system. Recent evidence supports a novel role for the immunoproteasome in the cellular stress response potentially through regulation of NFκB signaling, which is the primary response to multiple stressors. The current study tests whether the Classical or Alternative Pathways are regulated by immunoproteasome following chronic TNFα exposure in cultured retinal pigment epithelial cells isolated from wild-type mice and mice deficient in one (LMP2, L2) or two (LMP7 and MECL-1, L7M1) immunoproteasome subunits. Assays were performed to assess the expression of NFκB responsive genes, the content and activity of NFκB transcription factors (p65, p50, p52, cRel, RelB), and expression and content of regulatory proteins (IκBα, A20, RPS3). Major findings include distinct differences in expression of NFκB responsive genes in both KO cells. The mechanism responsible for the altered gene expression could not be established for L7M1 since no major differences in NFκB transcription factor content or activation were observed. However, L2 cells exhibited substantially higher content and diminished activation of NFκB transcription factors associated with the Alternative Pathway and delayed termination of the Classical Pathway. These results provide strong experimental evidence supporting a role for immunoproteasome in modulating NFκB signaling.

AB - Immunoproteasome is a protease abundant in immune cells and also present, albeit at lower concentrations, in cells outside the immune system. Recent evidence supports a novel role for the immunoproteasome in the cellular stress response potentially through regulation of NFκB signaling, which is the primary response to multiple stressors. The current study tests whether the Classical or Alternative Pathways are regulated by immunoproteasome following chronic TNFα exposure in cultured retinal pigment epithelial cells isolated from wild-type mice and mice deficient in one (LMP2, L2) or two (LMP7 and MECL-1, L7M1) immunoproteasome subunits. Assays were performed to assess the expression of NFκB responsive genes, the content and activity of NFκB transcription factors (p65, p50, p52, cRel, RelB), and expression and content of regulatory proteins (IκBα, A20, RPS3). Major findings include distinct differences in expression of NFκB responsive genes in both KO cells. The mechanism responsible for the altered gene expression could not be established for L7M1 since no major differences in NFκB transcription factor content or activation were observed. However, L2 cells exhibited substantially higher content and diminished activation of NFκB transcription factors associated with the Alternative Pathway and delayed termination of the Classical Pathway. These results provide strong experimental evidence supporting a role for immunoproteasome in modulating NFκB signaling.

UR - http://www.scopus.com/inward/record.url?scp=84874016734&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84874016734&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0056187

DO - 10.1371/journal.pone.0056187

M3 - Article

C2 - 23457524

AN - SCOPUS:84874016734

VL - 8

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 2

M1 - e56187

ER -