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Endoplazmik retikulum-aracılı protein yıkım yolağında görev yapan p97/VCP ve Ufd1-Npl4 adlı proteinlerin NF-κB yolağına etkisinin prostat kanser hücre hattında incelenmesi

Year 2018, , 142 - 146, 14.09.2018
https://doi.org/10.19161/etd.418150

Abstract

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
.

References

  • 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

Year 2018, , 142 - 146, 14.09.2018
https://doi.org/10.19161/etd.418150

Abstract

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
.

References

  • 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.
There are 17 citations in total.

Details

Primary Language Turkish
Subjects Health Care Administration
Journal Section Research Articles
Authors

Petek Ballar Kırmızıbayrak 0000-0002-6189-1818

Burcu Erbaykent Tepedelen 0000-0002-9565-6349

Publication Date September 14, 2018
Submission Date May 30, 2017
Published in Issue Year 2018

Cite

Vancouver Ballar Kırmızıbayrak P, Erbaykent Tepedelen B. Endoplazmik retikulum-aracılı protein yıkım yolağında görev yapan p97/VCP ve Ufd1-Npl4 adlı proteinlerin NF-κB yolağına etkisinin prostat kanser hücre hattında incelenmesi. ETD. 2018;57(3):142-6.

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