Endoplazmik retikulum-aracılı protein yıkım yolağında görev yapan p97/VCP ve Ufd1-Npl4 adlı proteinlerin NF-κB yolağına etkisinin prostat kanser hücre hattında incelenmesi
Öz
Amaç: Bu çalışmada, endoplazmik retikulum-aracılı yıkım yolağındaki
retrotranslokasyon basamağının anahtar proteini olan p97/VCP ve etkileşim
partnerleri olan Ufd1 ve Npl4 proteinlerinin NF-κB yolağı üzerine etkileri
prostat kanser hücre hattında incelendi.
Gereç ve
Yöntem: p97/VCP, Ufd1 ve Npl4 ifadeleri RNA interferans RNAi
(RNA interferans) teknolojisi ile LNCAP hücrelerinde susturuldu ve NF-κB
aktivitesi ikili lusiferaz yöntemi ile NF-κB yolağı proteinlerinin ifadesi ise
immünoblotlama ile değerlendirildi.
Bulgular: p97/VCP, Ufd1 ve Npl4 ifadelerinin susturulması LNCaP hücrelerinde NF-κB
aktivitesini anlamlı olarak azaltmış ve IκBα protein seviyesini arttırırken
fosforile NF-κB ve fosforile IκBα seviyelerini azaltmıştır.
Sonuç: Retrotranslokasyon kompleks üyelerinin susturulması ile LNCaP
hücrelerinde NF-κB aktivitesinin azalmasının NF-κB inhibitörü olan IκBα
seviyesinin artmasına bağlı olduğu düşünülmektedir. Bulgularımız p97/VCP ve
etkileşim partnerleri Ufd1-Npl4 proteinlerinin prostat kanserinde NF-κB
yolağının düzenlenmesinde önemli bir etken olabileceğini önermektedir.
Anahtar Kelimeler
Endoplazmik retikulum ilişkili yıkım yolağı retrotranslokasyon p97/VCP NF-κB prostat kanseri.
Kaynakça
- Ahner A, Brodsky JL. Checkpoints in ER-associated degradation: Excuse me, which way to the proteasome? Trends Cell Biol 2004;14(9):474-8.
- Meusser B, Hirsch C, Jarosch E, Sommer T. ERAD: The long road to destruction. Nat Cell Biol 2005;7(8):766-72.
- Hartl FU, Hayer-Hartl M. Converging concepts of protein folding in vitro and in vivo. Nat Struct Mol Biol 2009;16(6):574-81.
- Goldberg AL. Protein degradation and protection against misfolded or damaged proteins. Nature 2003;426(6968):895-9.
- Erzurumlu Y, Kose FA, Gozen O, Gozuacik D, Toth EA, Ballar P. A unique IBMPFD-related P97/VCP mutation with differential binding pattern and subcellular localization. Int J Biochem Cell Biol 2013;45(4):773-82.
- Fessart D, Marza E, Taouji S, Delom F, Chevet E. P97/CDC-48: Proteostasis control in tumor cell biology. Cancer Lett 2013;337(1):26-34.
- Magadán JG, Pérez-Victoria FJ, Sougrat R, Ye Y, Strebel K, Bonifacino JS. Multilayered mechanism of CD4 downregulation by HIV-1 Vpu involving distinct ER retention and ERAD targeting steps. PLoS Pathog 2010;6(4):e1000869.
- Kahn M, Sugawara H, McGowan P, et al. CD4+ T cell clones specific for the human p97 melanoma-associated antigen can eradicate pulmonary metastases from a murine tumor expressing the p97 antigen. J Immunol 1991;146(9):3235-41.
- Alexandru G, Graumann J, Smith GT, Kolawa NJ, Fang R, Deshaies RJ. UBXD7 binds multiple ubiquitin ligases and implicates p97 in HIF1a- turnover. Cell 2008;134(5):804-16.
- Hoffmann A, Baltimore D. Circuitry of nuclear factor kappa B signaling. Immunol Rev 2006;210(1):171-86.
- Israel A. The IKK complex, a central regulator of NF-kappaB activation. Cold Spring Harb Perspect Biol 2010;2(3): a000158.
- Erzurumlu Y, Ballar P. Androgen mediated regulation of endoplasmic reticulum-associated degradation and its effects on prostate cancer. Sci Rep 2017;16(7):40719.
- Asai T, Tomita Y, Nakatsuka S, et al. VCP (p97) regulates NFkappaB signaling pathway, which is important for metastasis of osteosarcoma cell line. Jpn J Cancer Res 2002;93(3):296-304.
- Lizano P, Rashed E, Kang H, et al. The valosin-containing protein promotes cardiac survival through the inducible isoform of nitric oxide synthase. Cardiovasc Res 2013;99(4):685-93.
- Li JM, Wu H, Zhang W, Blackburn MR, Jin J. The p97-UFD1L-NPL4 protein complex mediates cytokine-induced IκBα proteolysis. Mol Cell Biol 2014;34(3):335-47.
- Schweitzer K, Pralow A, Naumann M. p97/VCP promotes Cullin-RING-ubiquitin-ligase/proteasome-dependent degradation of IκBα and the preceding liberation of RelA from ubiquitinated IκBα. J Cell Mol Med 2016;20(1):58-70.
- Zhang Z, Wang Y, Li C, et al. The transitional endoplasmic reticulum ATPase p97 regulates the alternative nuclear factor NF-κB signaling via partial degradation of the NF-κB subunit p100. J Biol Chem 2015;290(32):19558-68.
Investigating the effect of p97/VCP and Ufd1-Npl4 proteins functioning in the endoplasmic reticulum-associated degradation on the NF-κB pathway
Öz
Aim: In this study, the effects of p97/VCP and its
interacting partners Ufd1 and Npl4 proteins, which function at the
retrotranslocation step of endoplasmic reticulum-associated degradation were
investigated on the NF-κB pathway in the prostate cancer cell line.
Materials and Methods: The expressions of p97/VCP,
Ufd1 and Npl4 were silenced in LNCaP cells using RNAi (RNA interference)
technology. NF-κB activity was evaluated by dual luciferase assay system and
the expression
of NF-κB pathway proteins were investigated using immunoblotting.
Results: Silencing of expression of p97/VCP, Ufd1 and Npl4
significantly diminished NF-κB activity and increased the level of IκBα protein
while decreased phosphorylated NF-κB and phosphorylated IκBα levels in LNCaP
cells.
Conclusion: The decrease NF-κB activity
upon silencing the expressions of retrotranslocation complex members in LNCaP
cells might be associated with the increased level of NF-κB inhibitor IκBα. Our
data suggests that p97/VCP and its interacting partners might be important
factors on the regulation of NF-κB pathway in the prostate cancer.
Anahtar Kelimeler
Endoplasmic reticulum-associated degradation retrotranslocation p97/VCP NF-κB prostate cancer.
Kaynakça
- Ahner A, Brodsky JL. Checkpoints in ER-associated degradation: Excuse me, which way to the proteasome? Trends Cell Biol 2004;14(9):474-8.
- Meusser B, Hirsch C, Jarosch E, Sommer T. ERAD: The long road to destruction. Nat Cell Biol 2005;7(8):766-72.
- Hartl FU, Hayer-Hartl M. Converging concepts of protein folding in vitro and in vivo. Nat Struct Mol Biol 2009;16(6):574-81.
- Goldberg AL. Protein degradation and protection against misfolded or damaged proteins. Nature 2003;426(6968):895-9.
- Erzurumlu Y, Kose FA, Gozen O, Gozuacik D, Toth EA, Ballar P. A unique IBMPFD-related P97/VCP mutation with differential binding pattern and subcellular localization. Int J Biochem Cell Biol 2013;45(4):773-82.
- Fessart D, Marza E, Taouji S, Delom F, Chevet E. P97/CDC-48: Proteostasis control in tumor cell biology. Cancer Lett 2013;337(1):26-34.
- Magadán JG, Pérez-Victoria FJ, Sougrat R, Ye Y, Strebel K, Bonifacino JS. Multilayered mechanism of CD4 downregulation by HIV-1 Vpu involving distinct ER retention and ERAD targeting steps. PLoS Pathog 2010;6(4):e1000869.
- Kahn M, Sugawara H, McGowan P, et al. CD4+ T cell clones specific for the human p97 melanoma-associated antigen can eradicate pulmonary metastases from a murine tumor expressing the p97 antigen. J Immunol 1991;146(9):3235-41.
- Alexandru G, Graumann J, Smith GT, Kolawa NJ, Fang R, Deshaies RJ. UBXD7 binds multiple ubiquitin ligases and implicates p97 in HIF1a- turnover. Cell 2008;134(5):804-16.
- Hoffmann A, Baltimore D. Circuitry of nuclear factor kappa B signaling. Immunol Rev 2006;210(1):171-86.
- Israel A. The IKK complex, a central regulator of NF-kappaB activation. Cold Spring Harb Perspect Biol 2010;2(3): a000158.
- Erzurumlu Y, Ballar P. Androgen mediated regulation of endoplasmic reticulum-associated degradation and its effects on prostate cancer. Sci Rep 2017;16(7):40719.
- Asai T, Tomita Y, Nakatsuka S, et al. VCP (p97) regulates NFkappaB signaling pathway, which is important for metastasis of osteosarcoma cell line. Jpn J Cancer Res 2002;93(3):296-304.
- Lizano P, Rashed E, Kang H, et al. The valosin-containing protein promotes cardiac survival through the inducible isoform of nitric oxide synthase. Cardiovasc Res 2013;99(4):685-93.
- Li JM, Wu H, Zhang W, Blackburn MR, Jin J. The p97-UFD1L-NPL4 protein complex mediates cytokine-induced IκBα proteolysis. Mol Cell Biol 2014;34(3):335-47.
- Schweitzer K, Pralow A, Naumann M. p97/VCP promotes Cullin-RING-ubiquitin-ligase/proteasome-dependent degradation of IκBα and the preceding liberation of RelA from ubiquitinated IκBα. J Cell Mol Med 2016;20(1):58-70.
- Zhang Z, Wang Y, Li C, et al. The transitional endoplasmic reticulum ATPase p97 regulates the alternative nuclear factor NF-κB signaling via partial degradation of the NF-κB subunit p100. J Biol Chem 2015;290(32):19558-68.
Ayrıntılar
| Birincil Dil | Türkçe |
|---|---|
| Konular | Sağlık Kurumları Yönetimi |
| Bölüm | Araştırma Makaleleri |
| Yazarlar | |
| Yayımlanma Tarihi | 14 Eylül 2018 |
| Gönderilme Tarihi | 30 Mayıs 2017 |
| Yayımlandığı Sayı | Yıl 2018 |