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The Assembly of EDC4 and Dcp1a into Processing Bodies Is Critical for the Translational Regulation of IL-6.

Seto E, Yoshida-Sugitani R, Kobayashi T, Toyama-Sorimachi N - PLoS ONE (2015)

Bottom Line: In contrast, P-body assembly in response to oxidative stress or TLR4 stimulation was increased in M1-THPs as compared to M2-THPs.These results suggest that mRNA metabolism is controlled differently in M1-THPs and M2-THPs.Interestingly, knocking down EDC4 or Dcp1a, which are components of P-bodies, severely reduced the production of IL-6, but not TNF-α in M1-THPs without decreasing the amount of IL-6 mRNA.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Immunology and Inflammation, Research Institute, National Center for Global Health and Medicine, Tokyo, Japan; Department of Virology and Preventive Medicine, Gunma University Graduate School of Medicine, Gunma, Japan.

ABSTRACT
Macrophages play critical roles in the onset of various diseases and in maintaining homeostasis. There are several functional subsets, of which M1 and M2 macrophages are of particular interest because they are differentially involved in inflammation and its resolution. Here, we investigated the differences in regulatory mechanisms between M1- and M2-polarized macrophages by examining mRNA metabolic machineries such as stress granules (SGs) and processing bodies (P-bodies). Human monocytic leukemia THP-1 cells cultured under M1-polarizing conditions (M1-THPs) had less ability to assemble oxidative-stress-induced SGs than those cultured under M2-polarizing conditions (M2-THPs). In contrast, P-body assembly in response to oxidative stress or TLR4 stimulation was increased in M1-THPs as compared to M2-THPs. These results suggest that mRNA metabolism is controlled differently in M1-THPs and M2-THPs. Interestingly, knocking down EDC4 or Dcp1a, which are components of P-bodies, severely reduced the production of IL-6, but not TNF-α in M1-THPs without decreasing the amount of IL-6 mRNA. This is the first report to demonstrate that the assembly of EDC4 and Dcp1a into P-bodies is critical in the posttranscriptional regulation of IL-6. Thus, improving our understanding of the mechanisms governing mRNA metabolism by examining macrophage subtypes may lead to new therapeutic targets.

No MeSH data available.


Related in: MedlinePlus

Oxidative stress does not induce P-body accumulation in M1-THPs or M2-THPs.(A) M1- and M2-polarized THP-1 cells, either untreated (NT) or treated with 0.5 mM arsenite (Ars) for 30 minutes, were fixed and stained with a goat polyclonal antibody specific for EDC4 and a mouse monoclonal antibody specific for Dcp1a, followed by staining with Alexa 488-conjugated donkey anti-goat IgG and Alexa 568-conjugated donkey anti-mouse secondary antibodies. (B) Bar graph showing the average number of EDC4- or Dcp1a- positive foci per cell. The fluorescent dots were counted as described in Methods, and the error bars indicate SD. **p<0.01, ***p<0.001
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pone.0123223.g002: Oxidative stress does not induce P-body accumulation in M1-THPs or M2-THPs.(A) M1- and M2-polarized THP-1 cells, either untreated (NT) or treated with 0.5 mM arsenite (Ars) for 30 minutes, were fixed and stained with a goat polyclonal antibody specific for EDC4 and a mouse monoclonal antibody specific for Dcp1a, followed by staining with Alexa 488-conjugated donkey anti-goat IgG and Alexa 568-conjugated donkey anti-mouse secondary antibodies. (B) Bar graph showing the average number of EDC4- or Dcp1a- positive foci per cell. The fluorescent dots were counted as described in Methods, and the error bars indicate SD. **p<0.01, ***p<0.001

Mentions: We next compared oxidative stress-induced P-body formation in M1-THPs and M2-THPs by staining with antibodies against Dcp1a or EDC4, key marker proteins that are linked to P-body formation[39,40]. EDC4-positive cytoplasmic foci were present in both M1-THPs and M2-THPs, and most of these were also positive for Dcp1a (Fig 2A). Although arsenite treatment did not increase the number of P-bodies in M1-THPs or M2-THPs, the M1-THPs were able to assemble more P-bodies than M2-THPs whether in the presence or absence of arsenite stimulation (Fig 2B). These results suggested that the regulation of P-body assembly differed in M1-THPs and M2-THPs.


The Assembly of EDC4 and Dcp1a into Processing Bodies Is Critical for the Translational Regulation of IL-6.

Seto E, Yoshida-Sugitani R, Kobayashi T, Toyama-Sorimachi N - PLoS ONE (2015)

Oxidative stress does not induce P-body accumulation in M1-THPs or M2-THPs.(A) M1- and M2-polarized THP-1 cells, either untreated (NT) or treated with 0.5 mM arsenite (Ars) for 30 minutes, were fixed and stained with a goat polyclonal antibody specific for EDC4 and a mouse monoclonal antibody specific for Dcp1a, followed by staining with Alexa 488-conjugated donkey anti-goat IgG and Alexa 568-conjugated donkey anti-mouse secondary antibodies. (B) Bar graph showing the average number of EDC4- or Dcp1a- positive foci per cell. The fluorescent dots were counted as described in Methods, and the error bars indicate SD. **p<0.01, ***p<0.001
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4430274&req=5

pone.0123223.g002: Oxidative stress does not induce P-body accumulation in M1-THPs or M2-THPs.(A) M1- and M2-polarized THP-1 cells, either untreated (NT) or treated with 0.5 mM arsenite (Ars) for 30 minutes, were fixed and stained with a goat polyclonal antibody specific for EDC4 and a mouse monoclonal antibody specific for Dcp1a, followed by staining with Alexa 488-conjugated donkey anti-goat IgG and Alexa 568-conjugated donkey anti-mouse secondary antibodies. (B) Bar graph showing the average number of EDC4- or Dcp1a- positive foci per cell. The fluorescent dots were counted as described in Methods, and the error bars indicate SD. **p<0.01, ***p<0.001
Mentions: We next compared oxidative stress-induced P-body formation in M1-THPs and M2-THPs by staining with antibodies against Dcp1a or EDC4, key marker proteins that are linked to P-body formation[39,40]. EDC4-positive cytoplasmic foci were present in both M1-THPs and M2-THPs, and most of these were also positive for Dcp1a (Fig 2A). Although arsenite treatment did not increase the number of P-bodies in M1-THPs or M2-THPs, the M1-THPs were able to assemble more P-bodies than M2-THPs whether in the presence or absence of arsenite stimulation (Fig 2B). These results suggested that the regulation of P-body assembly differed in M1-THPs and M2-THPs.

Bottom Line: In contrast, P-body assembly in response to oxidative stress or TLR4 stimulation was increased in M1-THPs as compared to M2-THPs.These results suggest that mRNA metabolism is controlled differently in M1-THPs and M2-THPs.Interestingly, knocking down EDC4 or Dcp1a, which are components of P-bodies, severely reduced the production of IL-6, but not TNF-α in M1-THPs without decreasing the amount of IL-6 mRNA.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Immunology and Inflammation, Research Institute, National Center for Global Health and Medicine, Tokyo, Japan; Department of Virology and Preventive Medicine, Gunma University Graduate School of Medicine, Gunma, Japan.

ABSTRACT
Macrophages play critical roles in the onset of various diseases and in maintaining homeostasis. There are several functional subsets, of which M1 and M2 macrophages are of particular interest because they are differentially involved in inflammation and its resolution. Here, we investigated the differences in regulatory mechanisms between M1- and M2-polarized macrophages by examining mRNA metabolic machineries such as stress granules (SGs) and processing bodies (P-bodies). Human monocytic leukemia THP-1 cells cultured under M1-polarizing conditions (M1-THPs) had less ability to assemble oxidative-stress-induced SGs than those cultured under M2-polarizing conditions (M2-THPs). In contrast, P-body assembly in response to oxidative stress or TLR4 stimulation was increased in M1-THPs as compared to M2-THPs. These results suggest that mRNA metabolism is controlled differently in M1-THPs and M2-THPs. Interestingly, knocking down EDC4 or Dcp1a, which are components of P-bodies, severely reduced the production of IL-6, but not TNF-α in M1-THPs without decreasing the amount of IL-6 mRNA. This is the first report to demonstrate that the assembly of EDC4 and Dcp1a into P-bodies is critical in the posttranscriptional regulation of IL-6. Thus, improving our understanding of the mechanisms governing mRNA metabolism by examining macrophage subtypes may lead to new therapeutic targets.

No MeSH data available.


Related in: MedlinePlus