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PLOS ONE  2011 

RPS3a Over-Expressed in HBV-Associated Hepatocellular Carcinoma Enhances the HBx-Induced NF-κB Signaling via Its Novel Chaperoning Function

DOI: 10.1371/journal.pone.0022258

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Abstract:

Hepatitis B virus (HBV) infection is one of the major causes of hepatocellular carcinoma (HCC) development. Hepatitis B virus X protein (HBx) is known to play a key role in the development of hepatocellular carcinoma (HCC). Several cellular proteins have been reported to be over-expressed in HBV-associated HCC tissues, but their role in the HBV-mediated oncogenesis remains largely unknown. Here, we explored the effect of the over-expressed cellular protein, a ribosomal protein S3a (RPS3a), on the HBx-induced NF-κB signaling as a critical step for HCC development. The enhancement of HBx-induced NF-κB signaling by RPS3a was investigated by its ability to translocate NF-κB (p65) into the nucleus and the knock-down analysis of RPS3a. Notably, further study revealed that the enhancement of NF-κB by RPS3a is mediated by its novel chaperoning activity toward physiological HBx. The over-expression of RPS3a significantly increased the solubility of highly aggregation-prone HBx. This chaperoning function of RPS3a for HBx is closely correlated with the enhanced NF-κB activity by RPS3a. In addition, the mutational study of RPS3a showed that its N-terminal domain (1–50 amino acids) is important for the chaperoning function and interaction with HBx. The results suggest that RPS3a, via extra-ribosomal chaperoning function for HBx, contributes to virally induced oncogenesis by enhancing HBx-induced NF-κB signaling pathway.

References

[1]  Branda M, Wands JR (2006) Signal transduction cascades and hepatitis B and C related hepatocellular carcinoma. Hepatology 43: 891–902.
[2]  Kremsdorf D, Soussan P, Paterlini-Brechot P, Brechot C (2006) Hepatitis B virus-related human carcinogenesis. Oncogene 25: 3823–3833.
[3]  Bouchard MJ, Schneider RJ (2004) The enigmatic X gene of hepatitis B virus. J Virol 78: 12725–12734.
[4]  Tang H, Oishi N, Kaneko S, Murakami S (2006) Molecular functions and biological roles of hepatitis B virus X protein. Cancer Sci 97: 977–983.
[5]  Wang WL, London WT, Lega L, Feitelson MA (1991) HBxAg in the liver from carrier patients with chronic hepatitis and cirrhosis. Hepatology 14: 29–37.
[6]  Benn J, Su F, Doria M, Schneider RJ (1996) Hepatitis B virus HBx protein induces transcription factor AP-1 by activation of extracellular signal-regulated and c-Jun N-terminal Mitogen-Activated Protein kinases. J Vriol 70: 4978–4985.
[7]  Karin M (2006) Nuclear factor-kappaB in cancer development and progression. Nature 441: 431–436.
[8]  Lucito R, Schneider RJ (1992) Hepatitis B virus protein activates transcription factor NF-κB without requirement for protein kinase C. J Virol 66: 983–991.
[9]  Wang T, Wang Y, Wu MC, Guan XY, Yin ZF (2004) Activating mechanism of transcriptor NF-kappaB regulated by hepatitis B virus X protein in hepatocellular carcinoma. World J Gastroenterol 10: 356–360.
[10]  Waris G, Huh KW, Siddiqui A (2001) Mitochondrially associated hepatitis B virus X protein constitutively activates transcription factors STAT-3 and NF-kappa B via oxidative stress. Mol Cell Biol 21: 7721–7730.
[11]  Yun C, Um HR, Jin YH, Wang JH, Lee MO, et al. (2002) NF-kappaB activation by hepatitis B virus X (HBx) protein shifts the cellular fate toward survival. Cancer Lett 184: 97–104.
[12]  Iizuka N, Oka M, Yamada-Okabe H, Mori N, Tamesa T, et al. (2002) Comparison of gene expression profiles between hepatitis B virus- and Hepatitis C virus-infected hepatocellular carcinoma by oligonucleotide microarray data on the basis of a supervised learning method. Cancer Res 62: 3939–3944.
[13]  Kim MY, Park E, Park JH, Park DH, Moon WS, et al. (2001) Expression profile of nine novel genes differentially expressed in hepatitis B virus-associated hepatocellular carcinomas. Oncogene 20: 4568–4575.
[14]  Kim JH, You KR, Kim IH, Cho BH, Kim CY, et al. (2004) Over-expression of the ribosomal protein L36a gene is associated with cellular proliferation in hepatocellular carcinoma. Hepatology 39: 129–138.
[15]  Takagi M, Absalon MJ, McLure KG, Kastan MB (2005) Regulation of p53 translation and induction after DNA damage by ribosomal protein L26 and Nucleodin. Cell 123: 49–53.
[16]  Lutsch G, Stahl J, K?rgel HJ, Noll F, Bielka H (1990) Immunoelectron microscopic studies on the location of ribosomal proteins on the surface of the 40S ribosomal subunit from rat liver. Eur J Cell Biol 51: 140–150.
[17]  Kashuba E, Yurchenko M, Szirak K, Stahl J, Klein G, et al. (2005) Epstein-Barr virus-encoded EBNA-5 binds to Epstein-Barr virus-induced Fte1/S3a protein. Exp Cell Res 303: 47–55.
[18]  Kho CJ, Wang Y, Zarbl H (1996) Effect of decreased fte-1 gene expression on protein synthesis, cell growth, and transformation. Cell Growth Differ 7: 1157–1166.
[19]  Kho CJ, Zarbl H (1992) Fte-1, a v-fos transformation effector gene, encodes the mammalian homologue of a yeast gene involved in protein import into mitochondria. Proc Natl Acad Sci U S A 89: 2200–2204.
[20]  Shuda M, Kondoh N, Tanaka K, Ryo A, Wakatsuki T, et al. (2000) Enhanced expression of translation factor mRNAs in hepatocellular carcinoma. Anticancer Res 20: 2489–2494.
[21]  Musholt TJ, Goodfellow PJ, Scheumann GF, Pichlmayr R, Wells SA Jr, et al. (1997) Differential display in primary and metastatic medullary thyroid carcinoma. J Surg Res 69: 94–100.
[22]  Pogue-geile K, Geiser JR, Shu M, Miller C, Wool IG, et al. (1991) Ribosomal protein Genes are overexpressed in colorectal cancer: isolation of a cDNA clone encoding the human S3 ribosomal protein. Mol Cell Biol 11: 3842–3849.
[23]  Slizhikova DK, Vinogradova TV, Sverdlov ED (2005) The NOLA2 and RPS3A genes as highly informative markers for human squamous cell lung cancer. Bioorg Khim 31: 195–199.
[24]  Starkey CR, Levy LS (1995) Identification of differentially expressed genes in T-lymphoid malignancies in an animal model system. Int J Cancer 62: 325–331.
[25]  Tarantul VZ, Nikolaev AI, Martynenko A, Hannig H, Hunsmann G, et al. (2000) Differential gene expression in B-cell non-Hodgkin's lymphoma of SIV-infected monkey. AIDS Res Hum Retroviruses 16: 173–179.
[26]  Naora H (1999) Involvement of ribosomal proteins in regulating cell growth and apoptosis: translational modulation or recruitment for extraribosomal activity? Immunol Cell Biol 77: 197–205.
[27]  Naora H, Takai I, Adachi M, Naora H (1998) Altered cellular responses by varying expression of a ribosomal protein gene: sequential coordination of enhancement and suppression of ribosomal protein S3a gene expression induces apoptosis. J Cell Biol 141: 741–753.
[28]  Naora H, Nishida T, Shindo Y, Adachi M, Naora H (1996) Constitutuvely enhanced nbl expression is associated with the induction of internucleosomal DNA cleavage by Actinomycin D. Biochem Biophys Res Commun 224: 258–264.
[29]  Song D, Sakamoto S, Taniguchi T (2002) Inhibition of Poly(ADP-ribose) polymerase activity by Bcl-2 in association with the ribosomal protein S3a. Biochemistry 41: 929–934.
[30]  Cui K, Coutts M, Stahl J, Sytkowski AJ (2000) Novel interaction between the transcription factor chop(GADD153) and the ribosomal protein FTE/S3a modulates erythropoiesis. J Biol Chem 275: 7591–7596.
[31]  Henkler F, Hoare J, Waseem N, Goldin RD, McGarvey MJ, et al. (2001) Intracellular localization of the hepatitis B virus HBx protein. J Gen Virol 82: 871–882.
[32]  Kim KH, Seong BL (2003) Pro-apoptotic function of HBV X protein is mediated by interaction with c-FLIP and enhancement of death-inducing signal. EMBO J 22: 2104–2116.
[33]  Song CZ, Bai ZL, Song CC, Wang QW (2003) Aggregate formation of hepatitis B virus X protein affects cell cycle and apoptosis. World J Gastroenterol 9: 1521–1524.
[34]  Urban S, Hildt E, Eckerskorn C, Sirma H, Kekule A, et al. (1997) Isolation and molecular characterization of hepatitis B virsus X-protein from a baculovirus expression system. Hepatology 26: 1045–1053.
[35]  Jameel S, Siddiqui A, Maguire HF, Rao KV (1990) Hepatitis B virus X protein produced in Escherichia coli is biologically functional. J Virol 64: 3963–3966.
[36]  Liu D, Zou L, Li W, Wang L, Wu Y (2009) High-level expression and large-scale preparation of soluble HBx antigen from Escherichia coli. Biotechnol Appl Biochem 54: 141–147.
[37]  Marczinovits I, Somogyi C, Patthy A, Nemeth P, Molnar J (1997) An alternative purification protocol for producing hepatitis B Virus X antigen on a preparative scale in Escherichia coli. J Biotechnol 56: 81–88.
[38]  Calderwood SK, Khaleque MA, Sawyer DB, Ciocca DR (2006) Heat shock proteins in cancer: chaperones of tumorigenesis. Trends Biochem Sci 31: 164–172.
[39]  Tsutsumi S, Neckers L (2007) Extracellular heat shock protein 90: A role for a molecule chaperone in cell motility and cancer metstasis. Cancer Sci 98: 1536–1539.
[40]  Whitesell L, Lindquist SL (2005) HSP90 and the chaperoning of cancer. Nat Rev Cancer 5: 761–772.
[41]  Kovacs D, Rakacs M, Agoston B, Lenkey K, Semrad K, et al. (2009) Janus chaperones: Assistance of both RNA- and protein-folding by ribosomal proteins. FEBS Lett 583: 88–92.
[42]  Kabir MA, Sherman F (2008) Overexpressed ribosomal proteins suppress defective chaperonins in Saccharomyces cerevisiae. FEMS Yeast Res 8: 1236–1244.
[43]  Choi SI, Ryu K, Seong BL (2009) RNA-mediated chaperone type for de novo protein folding. RNA Biol 6: 21–24.
[44]  Kim HK, Choi SI, Seong BL (2010) 5S rRNA-assisted DnaK refolding. Biochem Biophys Res Commun 391: 1177–1181.
[45]  Choi SI, Han KS, Kim CW, Ryu K, Kim BH, et al. (2008) Protein solubility and folding enhancement by interaction with RNA. PLoS One 3: e2677.
[46]  Kim CW, Han KS, Ryu KS, Kim BH, Kim KH, et al. (2007) N-terminal domains of native multidomain proteins have the potential to assist de novo folding of their downstream domains in vivo by acting as solubility enhancers. Protein Sci 16: 635–643.
[47]  Trepel J, Mollapour M, Giaccone G, Neckers L (2010) Targeting the dynamic HSP90 compelex in cancer. Nat Rev Cancer 10: 537–549.

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