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The E3 ubiquitin ligase seven in absentia homolog 1 may be a potential new therapeutic target for Parkinson's disease.

Cai ZL, Xu J, Xue SR, Liu YY, Zhang YJ, Zhang XZ, Wang X, Wu FP, Li XM - Neural Regen Res (2015)

Bottom Line: In this study, we investigated the effect of an antibody against E3 ubiquitin ligase seven in absentia homolog 1 (SIAH-1) in PC12 cells. 1-Methyl-4-phenylpyridinium (MPP(+)) treatment increased α-synuclein, E1 and SIAH-1 protein levels in PC12 cells, and it reduced cell viability; however, there was no significant change in light chain 3 expression.Combined treatment with MPP(+) and rapamycin reduced SIAH-1 and α-synuclein levels.These findings suggest that the SIAH-1 antibody reduces the monoubiquitination and aggregation of α-synuclein, promoting its degradation by the ubiquitin-proteasome pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Affiliated Lianyungang Hospital of Xuzhou Medical College, Lianyungang, Jiangsu Province, China.

ABSTRACT
In this study, we investigated the effect of an antibody against E3 ubiquitin ligase seven in absentia homolog 1 (SIAH-1) in PC12 cells. 1-Methyl-4-phenylpyridinium (MPP(+)) treatment increased α-synuclein, E1 and SIAH-1 protein levels in PC12 cells, and it reduced cell viability; however, there was no significant change in light chain 3 expression. Treatment with an SIAH-1 antibody decreased mRNA expression levels of α-synuclein, light chain 3 and SIAH-1, but increased E1 mRNA expression. It also increased cell viability. Combined treatment with MPP(+) and rapamycin reduced SIAH-1 and α-synuclein levels. Treatment with SIAH-1 antibody alone diminished α-synuclein immunoreactivity in PC12 cells, and reduced the colocalization of α-synuclein and light chain 3. These findings suggest that the SIAH-1 antibody reduces the monoubiquitination and aggregation of α-synuclein, promoting its degradation by the ubiquitin-proteasome pathway. Consequently, SIAH-1 may be a potential new therapeutic target for Parkinson's disease.

No MeSH data available.


Related in: MedlinePlus

Effects of SIAH-1 antibody treatment on α-synuclein and the autophagy degradation pathway in PC12 cells.Western blot assay for protein levels (upper panel) and statistical analysis of optical density measurements (target protein/β-actin; lower panel) in PC12 cells after treatment with MPP+, RAPA and SIAH-1 antibody. (A) SIAH-1 (37 kDa); (B) α-synuclein (19 kDa); (C) p53 (55 kDa); (D) LC3-II (17 kDa); (E) E1 (117 kDa). Values are represented as the mean ± SD (n = 5) (one-way analysis of variance followed by Fisher's least significant difference post hoc test). #P < 0.05, ##P < 0.01, vs. control group (MPP+, RAPA and anti-SIAH-1 are “–”). **P < 0.01, vs. MPP+ group (MPP+ is “+”; RAPA and anti-SIAH-1 are “–”). SIAH-1: E3 ubiquitin ligase seven in absentia homolog 1; LC3: light chain 3; MPP+: 1-methyl-4-phenylpyridinium; RAPA: rapamycin.
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Figure 2: Effects of SIAH-1 antibody treatment on α-synuclein and the autophagy degradation pathway in PC12 cells.Western blot assay for protein levels (upper panel) and statistical analysis of optical density measurements (target protein/β-actin; lower panel) in PC12 cells after treatment with MPP+, RAPA and SIAH-1 antibody. (A) SIAH-1 (37 kDa); (B) α-synuclein (19 kDa); (C) p53 (55 kDa); (D) LC3-II (17 kDa); (E) E1 (117 kDa). Values are represented as the mean ± SD (n = 5) (one-way analysis of variance followed by Fisher's least significant difference post hoc test). #P < 0.05, ##P < 0.01, vs. control group (MPP+, RAPA and anti-SIAH-1 are “–”). **P < 0.01, vs. MPP+ group (MPP+ is “+”; RAPA and anti-SIAH-1 are “–”). SIAH-1: E3 ubiquitin ligase seven in absentia homolog 1; LC3: light chain 3; MPP+: 1-methyl-4-phenylpyridinium; RAPA: rapamycin.

Mentions: MPP+ treatment not only increased the expression of α-synuclein (P < 0.01) and E1 (P < 0.01), but also that of SIAH-1 (P < 0.01). We found a nonsignificant increase in LC3-II expression (P = 0.17) (Figure 2). In addition, we observed significant α-synuclein aggregation and localization in the cytoplasm after MPP+ treatment. SIAH-1 immunofluorescence staining was significantly increased after 24 hours of MPP+ treatment (Figure 3).


The E3 ubiquitin ligase seven in absentia homolog 1 may be a potential new therapeutic target for Parkinson's disease.

Cai ZL, Xu J, Xue SR, Liu YY, Zhang YJ, Zhang XZ, Wang X, Wu FP, Li XM - Neural Regen Res (2015)

Effects of SIAH-1 antibody treatment on α-synuclein and the autophagy degradation pathway in PC12 cells.Western blot assay for protein levels (upper panel) and statistical analysis of optical density measurements (target protein/β-actin; lower panel) in PC12 cells after treatment with MPP+, RAPA and SIAH-1 antibody. (A) SIAH-1 (37 kDa); (B) α-synuclein (19 kDa); (C) p53 (55 kDa); (D) LC3-II (17 kDa); (E) E1 (117 kDa). Values are represented as the mean ± SD (n = 5) (one-way analysis of variance followed by Fisher's least significant difference post hoc test). #P < 0.05, ##P < 0.01, vs. control group (MPP+, RAPA and anti-SIAH-1 are “–”). **P < 0.01, vs. MPP+ group (MPP+ is “+”; RAPA and anti-SIAH-1 are “–”). SIAH-1: E3 ubiquitin ligase seven in absentia homolog 1; LC3: light chain 3; MPP+: 1-methyl-4-phenylpyridinium; RAPA: rapamycin.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
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Figure 2: Effects of SIAH-1 antibody treatment on α-synuclein and the autophagy degradation pathway in PC12 cells.Western blot assay for protein levels (upper panel) and statistical analysis of optical density measurements (target protein/β-actin; lower panel) in PC12 cells after treatment with MPP+, RAPA and SIAH-1 antibody. (A) SIAH-1 (37 kDa); (B) α-synuclein (19 kDa); (C) p53 (55 kDa); (D) LC3-II (17 kDa); (E) E1 (117 kDa). Values are represented as the mean ± SD (n = 5) (one-way analysis of variance followed by Fisher's least significant difference post hoc test). #P < 0.05, ##P < 0.01, vs. control group (MPP+, RAPA and anti-SIAH-1 are “–”). **P < 0.01, vs. MPP+ group (MPP+ is “+”; RAPA and anti-SIAH-1 are “–”). SIAH-1: E3 ubiquitin ligase seven in absentia homolog 1; LC3: light chain 3; MPP+: 1-methyl-4-phenylpyridinium; RAPA: rapamycin.
Mentions: MPP+ treatment not only increased the expression of α-synuclein (P < 0.01) and E1 (P < 0.01), but also that of SIAH-1 (P < 0.01). We found a nonsignificant increase in LC3-II expression (P = 0.17) (Figure 2). In addition, we observed significant α-synuclein aggregation and localization in the cytoplasm after MPP+ treatment. SIAH-1 immunofluorescence staining was significantly increased after 24 hours of MPP+ treatment (Figure 3).

Bottom Line: In this study, we investigated the effect of an antibody against E3 ubiquitin ligase seven in absentia homolog 1 (SIAH-1) in PC12 cells. 1-Methyl-4-phenylpyridinium (MPP(+)) treatment increased α-synuclein, E1 and SIAH-1 protein levels in PC12 cells, and it reduced cell viability; however, there was no significant change in light chain 3 expression.Combined treatment with MPP(+) and rapamycin reduced SIAH-1 and α-synuclein levels.These findings suggest that the SIAH-1 antibody reduces the monoubiquitination and aggregation of α-synuclein, promoting its degradation by the ubiquitin-proteasome pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Affiliated Lianyungang Hospital of Xuzhou Medical College, Lianyungang, Jiangsu Province, China.

ABSTRACT
In this study, we investigated the effect of an antibody against E3 ubiquitin ligase seven in absentia homolog 1 (SIAH-1) in PC12 cells. 1-Methyl-4-phenylpyridinium (MPP(+)) treatment increased α-synuclein, E1 and SIAH-1 protein levels in PC12 cells, and it reduced cell viability; however, there was no significant change in light chain 3 expression. Treatment with an SIAH-1 antibody decreased mRNA expression levels of α-synuclein, light chain 3 and SIAH-1, but increased E1 mRNA expression. It also increased cell viability. Combined treatment with MPP(+) and rapamycin reduced SIAH-1 and α-synuclein levels. Treatment with SIAH-1 antibody alone diminished α-synuclein immunoreactivity in PC12 cells, and reduced the colocalization of α-synuclein and light chain 3. These findings suggest that the SIAH-1 antibody reduces the monoubiquitination and aggregation of α-synuclein, promoting its degradation by the ubiquitin-proteasome pathway. Consequently, SIAH-1 may be a potential new therapeutic target for Parkinson's disease.

No MeSH data available.


Related in: MedlinePlus