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p300-mediated acetylation of COMMD1 regulates its stability, and the ubiquitylation and nucleolar translocation of the RelA NF-κB subunit.

O'Hara A, Simpson J, Morin P, Loveridge CJ, Williams AC, Novo SM, Stark LA - J. Cell. Sci. (2014)

Bottom Line: We show that p300 is required for stress-mediated ubiquitylation and nucleolar translocation of RelA, but that this effect is indirect.In contrast, tumour necrosis factor (TNF) has no effect on COMMD1 acetylation.Finally, we demonstrate these findings have relevance in a whole tissue setting.

View Article: PubMed Central - PubMed

Affiliation: Edinburgh Cancer Research Centre, IGMM, University of Edinburgh, Western General Hospital, Crewe Road, Edinburgh EH4 2XU, UK.

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COMMD1 acetylation and deacetylation protects against XIAP-mediated degradation. (A) SW480 cells were transfected with control, p300 and XIAP siRNA, either individually or together. Western blotting (WB) was performed on whole cell lysates. Actin controlled for loading. COMMD1 levels are depleted by p300 knockdown but rescued by concomitant knockdown of XIAP. (B) Cells were transfected with GST–COMMD1 alongside control or XIAP siRNA. COMMD1 was isolated and the level of acetylated (Ac) COMMD1 was determined as in Fig. 2. Input levels of XIAP confirm knockdown. (C) Cells were not treated (NT) or treated with aspirin (5 mM) or trichostatin A (TSA, 400 nM) for 16 h. The western blot shows COMMD1 and XIAP levels.
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f03: COMMD1 acetylation and deacetylation protects against XIAP-mediated degradation. (A) SW480 cells were transfected with control, p300 and XIAP siRNA, either individually or together. Western blotting (WB) was performed on whole cell lysates. Actin controlled for loading. COMMD1 levels are depleted by p300 knockdown but rescued by concomitant knockdown of XIAP. (B) Cells were transfected with GST–COMMD1 alongside control or XIAP siRNA. COMMD1 was isolated and the level of acetylated (Ac) COMMD1 was determined as in Fig. 2. Input levels of XIAP confirm knockdown. (C) Cells were not treated (NT) or treated with aspirin (5 mM) or trichostatin A (TSA, 400 nM) for 16 h. The western blot shows COMMD1 and XIAP levels.

Mentions: Previous studies have indicated COMMD1 is ubiquitylated by the E3 ligase XIAP, then targeted for degradation (Mufti et al., 2007). Therefore, we next determined whether this pathway was involved in COMMD1 regulation by p300. Fig. 3A demonstrates that the reduction in COMMD1 observed upon loss of p300 is reversed by concomitant depletion of XIAP.


p300-mediated acetylation of COMMD1 regulates its stability, and the ubiquitylation and nucleolar translocation of the RelA NF-κB subunit.

O'Hara A, Simpson J, Morin P, Loveridge CJ, Williams AC, Novo SM, Stark LA - J. Cell. Sci. (2014)

COMMD1 acetylation and deacetylation protects against XIAP-mediated degradation. (A) SW480 cells were transfected with control, p300 and XIAP siRNA, either individually or together. Western blotting (WB) was performed on whole cell lysates. Actin controlled for loading. COMMD1 levels are depleted by p300 knockdown but rescued by concomitant knockdown of XIAP. (B) Cells were transfected with GST–COMMD1 alongside control or XIAP siRNA. COMMD1 was isolated and the level of acetylated (Ac) COMMD1 was determined as in Fig. 2. Input levels of XIAP confirm knockdown. (C) Cells were not treated (NT) or treated with aspirin (5 mM) or trichostatin A (TSA, 400 nM) for 16 h. The western blot shows COMMD1 and XIAP levels.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150058&req=5

f03: COMMD1 acetylation and deacetylation protects against XIAP-mediated degradation. (A) SW480 cells were transfected with control, p300 and XIAP siRNA, either individually or together. Western blotting (WB) was performed on whole cell lysates. Actin controlled for loading. COMMD1 levels are depleted by p300 knockdown but rescued by concomitant knockdown of XIAP. (B) Cells were transfected with GST–COMMD1 alongside control or XIAP siRNA. COMMD1 was isolated and the level of acetylated (Ac) COMMD1 was determined as in Fig. 2. Input levels of XIAP confirm knockdown. (C) Cells were not treated (NT) or treated with aspirin (5 mM) or trichostatin A (TSA, 400 nM) for 16 h. The western blot shows COMMD1 and XIAP levels.
Mentions: Previous studies have indicated COMMD1 is ubiquitylated by the E3 ligase XIAP, then targeted for degradation (Mufti et al., 2007). Therefore, we next determined whether this pathway was involved in COMMD1 regulation by p300. Fig. 3A demonstrates that the reduction in COMMD1 observed upon loss of p300 is reversed by concomitant depletion of XIAP.

Bottom Line: We show that p300 is required for stress-mediated ubiquitylation and nucleolar translocation of RelA, but that this effect is indirect.In contrast, tumour necrosis factor (TNF) has no effect on COMMD1 acetylation.Finally, we demonstrate these findings have relevance in a whole tissue setting.

View Article: PubMed Central - PubMed

Affiliation: Edinburgh Cancer Research Centre, IGMM, University of Edinburgh, Western General Hospital, Crewe Road, Edinburgh EH4 2XU, UK.

Show MeSH
Related in: MedlinePlus