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Proteasome Composition in Cytokine-Treated Neurons and Astrocytes is Determined Mainly by Subunit Displacement

  • Kara L. Shanley
  • Che-Lin Hu
  • Oscar A. BizzozeroEmail author
Original Paper
  • 33 Downloads

Abstract

In this study, we investigated if subunit displacement and/or alterations in proteasome biosynthesis are responsible for the changes in the levels of constitutive proteasomes (c-20S), immunoproteasomes (i-20S) and the activators PA28 and PA700 in neurons and astrocytes cultured with a cytokine mixture (IFN-γ/TNF-α/IL-1β). Exposure of both cell types to cytokines for 24 h increases mRNA and protein expression of the i-20S-specific subunit β5i and PA28α/β, and leads to a decline in the amount of the c-20S-specific subunit β5. Since β5 mRNA levels are unchanged by the cytokine treatment, it is fair to conclude that displacement of constitutive β-subunits with inducible β5i subunits is likely the mechanism underlying the decrease in c-20S. As expected, the increase in the amount of the IFN-γ-inducible subunits coincides with elevated expression of phospho-STAT-1 and interferon regulatory factor-1 (IRF-1). However, inhibition of NF-κB signaling in cytokine-treated astrocytes reduces IRF-1 expression without affecting that of i-20S, c-20S and PA28. This suggests that STAT-1 is capable of increasing the transcription of i20S-specific subunits and PA28α/β by itself. The lack of a decrease in proteasome β5 mRNA expression is consistent with the fact that Nrf1 (Nfe2l1) and Nrf2 (Nfe2l2) levels are not reduced by pro-inflammatory cytokines. In contrast, we previously found that there is a significant Nrf1 dysregulation and reduced β5 mRNA expression in the spinal cords of mice with experimental autoimmune encephalomyelitis (EAE). Thus, there are stressors in EAE, other than a pro-inflammatory environment, that are not present in cytokine-treated cells.

Keywords

Astrocytes Cytokines Immunoproteasomes Neurons Nfe2l1 Nfe2l2 PA28 Proteasomes 

Abbreviations

α7 (Psma7)

20S subunit α7

β5 (Psmb5)

C-20S subunit β5

β5i (Psmb8)

I-20S inducible subunit β5

c-20S

Constitutive-20S particle

EAE

Experimental autoimmune encephalomyelitis

H3

Histone H3

HDAC2

Histone deacetylase 2

i-20S

Immuno-20S particle

IL-1β

Interleukin-1β

IFN-γ

Interferon-γ

IRF-1

Interferon regulatory factor-1

mTOR

Mammalian target of rapamycin

Nrf1 (Nfe2l1)

Nuclear factor (erythroid-derived 2)-like 1

Nrf2 (Nfe2l2)

Nuclear factor (erythroid-derived 2)-like 2

NF-κB

Nuclear factor kappa-light-chain-enhancer of activated B cells

PA28

11S regulatory particle

PA28α (Psme1)

PA28 subunit α

PA28β (Psme2)

PA28 subunit β

PA700

19S regulatory particle

PBX1

Pre-B-cell leukemia homeobox 1

qPCR

Quantitative polymerase chain reaction

Rpt5 (Psmc3)

19S regulatory subunit 6A

STAT-1

Signal transducer and activator of transcription 1

TNF-α

Tumor necrosis factor-α

Notes

Acknowledgements

This work was supported by PHHS Grants NS082805 from the National Institutes of Health.

Funding

Funding was provided by National Institute of Neurological Disorders and Stroke (Grant No. NS082805).

Supplementary material

11064_2020_2958_MOESM1_ESM.pdf (650 kb)
Supplementary file1 (PDF 650 kb)

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Authors and Affiliations

  1. 1.Department of Cell Biology and PhysiologyUniversity of New Mexico - Health Sciences CenterAlbuquerqueUSA
  2. 2.Department of Cell Biology and PhysiologyUniversity of New Mexico School of MedicineAlbuquerqueUSA

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