Archives of Virology

, Volume 164, Issue 5, pp 1271–1285 | Cite as

Hepatitis C virus core protein interacts with cellular metastasis suppressor Nm23-H1 and promotes cell migration and invasion

  • Catherine Paul
  • Lohit Khera
  • Rajeev KaulEmail author
Original Article


Hepatitis C virus (HCV) is the major etiological agent of hepatocellular carcinoma (HCC), which is the fourth most common cause of cancer-related deaths worldwide and second in terms of deaths of males (Bray et al. in CA Cancer J Clin 68(6):394–424, 2018). HCV-induced HCC is a multi-step process that involves alteration of several host regulatory pathways. One of the key features of HCV-associated hepatocellular carcinoma is the metastasis of cancer cells to different organs. Human Nm23-H1 is one of the best-studied metastasis suppressor proteins, and it has been shown to be modulated in many human cancers. Our study shows that the core protein of HCV genotype 2a can co-localize and interact directly with Nm23-H1 within cancer cells, resulting in modulation of the anti-metastasis properties of Nm23-H1. The HCV core protein promotes SUMOylation and degradation of the Nm23-H1 protein, as well as transcriptional downregulation. This study provides evidence that the HCV core protein is a pro-metastatic protein that can interact directly with and modulate the functions of cellular metastasis suppressor Nm23-H1.



This work was supported by Grants from DBT, Ministry of Science and Technology (BT/PR15109/GBD/27/320/2011), a PURSE Grant from DST, and an R&D Grant from the University of Delhi. CP and LK received a Senior Research Fellowship funded by CSIR (09/045(1242)/2012-EMR-1 and 09/045(1135)/2011-EMR-1). This work also used original research material provided by Apath LLC for academic use via an ‘Academic Material Transfer Agreement’.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

705_2019_4151_MOESM1_ESM.pdf (582 kb)
Fig. S1 HCV Core expression in 293T cells promotes cancer cell migration and invasion by overcoming effect of metastasis suppressor Nm23-H1. 293T cells expressing the HCV core protein show significantly increased migration as compared to control cells. Overexpression of the HCV core protein can overcome the effect of metastasis suppressor Nm23-H1. Fig. S2 HCV Core expression in Huh7.5 cells promotes cancer cell migration and invasion by overcoming effect of metastasis suppressor Nm23-H1. Huh7.5 cells expressing the HCV core protein show significantly increased migration as compared to control cells. Overexpression of the HCV core protein can overcome the effect of metastasis suppressor Nm23-H1. (PDF 582 kb)


  1. 1.
    Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A (2018) Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 68(6):394–424. CrossRefGoogle Scholar
  2. 2.
    Tang H, Grise H (2009) Cellular and molecular biology of HCV infection and hepatitis. Clin Sci (Lond) 117(2):49–65. CrossRefGoogle Scholar
  3. 3.
    Mohamed AA, Elbedewy TA, El-Serafy M, El-Toukhy N, Ahmed W, Ali El Din Z (2015) Hepatitis C virus: a global view. World J Hepatol 7:2676–2680. CrossRefGoogle Scholar
  4. 4.
    Grakoui A, Wychowski C, Lin C, Feinstone SM, Rice CM (1993) Expression and identification of hepatitis C virus polyprotein cleavage products. J Virol 67(3):1385–1395Google Scholar
  5. 5.
    Santolini E, Migliaccio G, La Monica N (1994) Biosynthesis and biochemical properties of the hepatitis C virus core protein. J Virol 68(6):3631–3641Google Scholar
  6. 6.
    Suzuki R, Matsuura Y, Suzuki T, Ando A, Chiba J, Harada S, Saito I, Miyamura T (1995) Nuclear localization of the truncated hepatitis C virus core protein with its hydrophobic C terminus deleted. J Gen Virol 76(Pt 1):53–61. CrossRefGoogle Scholar
  7. 7.
    Chang SC, Yen JH, Kang HY, Jang MH, Chang MF (1994) Nuclear localization signals in the core protein of hepatitis C virus. Biochem Biophys Res Commun 205(2):1284–1290. CrossRefGoogle Scholar
  8. 8.
    Suzuki R, Sakamoto S, Tsutsumi T, Rikimaru A, Tanaka K, Shimoike T, Moriishi K, Iwasaki T, Mizumoto K, Matsuura Y, Miyamura T, Suzuki T (2005) Molecular determinants for subcellular localization of hepatitis C virus core protein. J Virol 79(2):1271–1281. CrossRefGoogle Scholar
  9. 9.
    Nolandt O, Kern V, Muller H, Pfaff E, Theilmann L, Welker R, Krausslich HG (1997) Analysis of hepatitis C virus core protein interaction domains. J Gen Virol 78(Pt 6):1331–1340. CrossRefGoogle Scholar
  10. 10.
    Majeau N, Gagne V, Boivin A, Bolduc M, Majeau JA, Ouellet D, Leclerc D (2004) The N-terminal half of the core protein of hepatitis C virus is sufficient for nucleocapsid formation. J Gen Virol 85(Pt 4):971–981. CrossRefGoogle Scholar
  11. 11.
    Klein KC, Polyak SJ, Lingappa JR (2004) Unique features of hepatitis C virus capsid formation revealed by de novo cell-free assembly. J Virol 78(17):9257–9269. CrossRefGoogle Scholar
  12. 12.
    McLauchlan J (2000) Properties of the hepatitis C virus core protein: a structural protein that modulates cellular processes. J Viral Hepat 7(1):2–14CrossRefGoogle Scholar
  13. 13.
    Chaffer CL, Weinberg RA (2011) A perspective on cancer cell metastasis. Science 331(6024):1559–1564. CrossRefGoogle Scholar
  14. 14.
    Hartsough MT, Steeg PS (1998) Nm23-H1: genetic alterations and expression patterns in tumor metastasis. Am J Hum Genet 63(1):6–10. CrossRefGoogle Scholar
  15. 15.
    You DJ, Park CR, Lee HB, Moon MJ, Kang JH, Lee C, Oh SH, Ahn C, Seong JY, Hwang JI (2014) A splicing variant of NME1 negatively regulates NF-kappaB signaling and inhibits cancer metastasis by interacting with IKKbeta. J Biol Chem 289(25):17709–17720. CrossRefGoogle Scholar
  16. 16.
    Ma D, McCorkle JR, Kaetzel DM (2004) The metastasis suppressor NM23-H1 possesses 3′–5′ exonuclease activity. J Biol Chem 279(17):18073–18084. CrossRefGoogle Scholar
  17. 17.
    Hartsough MT, Morrison DK, Salerno M, Palmieri D, Ouatas T, Mair M, Patrick J, Steeg PS (2002) Nm23-H1 metastasis suppressor phosphorylation of kinase suppressor of Ras via a histidine protein kinase pathway. J Biol Chem 277(35):32389–32399. CrossRefGoogle Scholar
  18. 18.
    Kim HD, Youn B, Kim TS, Kim SH, Shin HS, Kim J (2009) Regulators affecting the metastasis suppressor activity of Nm23-H1. Mol Cell Biochem 329(1–2):167–173. CrossRefGoogle Scholar
  19. 19.
    Mileo AM, Piombino E, Severino A, Tritarelli A, Paggi MG, Lombardi D (2006) Multiple interference of the human papillomavirus-16 E7 oncoprotein with the functional role of the metastasis suppressor Nm23-H1 protein. J Bioenerg Biomembr 38(3–4):215–225. CrossRefGoogle Scholar
  20. 20.
    Tan CY, Chang CL (2018) NDPKA is not just a metastasis suppressor—be aware of its metastasis-promoting role in neuroblastoma. Lab Investig 98(2):219–227. CrossRefGoogle Scholar
  21. 21.
    MacDonald NJ, Freije JM, Stracke ML, Manrow RE, Steeg PS (1996) Site-directed mutagenesis of nm23-H1. Mutation of proline 96 or serine 120 abrogates its motility inhibitory activity upon transfection into human breast carcinoma cells. J Biol Chem 271(41):25107–25116CrossRefGoogle Scholar
  22. 22.
    Zhou Q, Yang X, Zhu D, Ma L, Zhu W, Sun Z, Yang Q (2007) Double mutant P96S/S120G of Nm23-H1 abrogates its NDPK activity and motility-suppressive ability. Biochem Biophys Res Commun 356(2):348–353. CrossRefGoogle Scholar
  23. 23.
    Miyagawa S, Kawasaki S, Makuuchi M (1996) Comparison of the characteristics of hepatocellular carcinoma between hepatitis B and C viral infection: tumor multicentricity in cirrhotic liver with hepatitis C. Hepatology 24(2):307–310. CrossRefGoogle Scholar
  24. 24.
    Tang ZY (2001) Hepatocellular carcinoma—cause, treatment and metastasis. World J Gastroenterol 7(4):445–454CrossRefGoogle Scholar
  25. 25.
    Trinchet JC, Ganne-Carrie N, Nahon P, N’Kontchou G, Beaugrand M (2007) Hepatocellular carcinoma in patients with hepatitis C virus-related chronic liver disease. World J Gastroenterol 13(17):2455–2460CrossRefGoogle Scholar
  26. 26.
    Khera L, Paul C, Kaul R (2017) Hepatitis C Virus E1 protein promotes cell migration and invasion by modulating cellular metastasis suppressor Nm23-H1. Virology 506:110–120. CrossRefGoogle Scholar
  27. 27.
    Fancsalszky L, Monostori E, Farkas Z, Pourkarimi E, Masoudi N, Hargitai B, Bosnar MH, Dezeljin M, Zsakai A, Vellai T, Mehta A, Takacs-Vellai K (2014) NDK-1, the homolog of NM23-H1/H2 regulates cell migration and apoptotic engulfment in C. elegans. PLoS One 9(3):e92687. CrossRefGoogle Scholar
  28. 28.
    Wakita T, Pietschmann T, Kato T, Date T, Miyamoto M, Zhao Z, Murthy K, Habermann A, Krausslich HG, Mizokami M, Bartenschlager R, Liang TJ (2005) Production of infectious hepatitis C virus in tissue culture from a cloned viral genome. Nat Med 11(7):791–796. CrossRefGoogle Scholar
  29. 29.
    Rueden CT, Schindelin J, Hiner MC, DeZonia BE, Walter AE, Arena ET, Eliceiri KW (2017) Image J2: ImageJ for the next generation of scientific image data. BMC Bioinform 18(1):529. CrossRefGoogle Scholar
  30. 30.
    Kaul R, Murakami M, Lan K, Choudhuri T, Robertson ES (2009) EBNA3C can modulate the activities of the transcription factor Necdin in association with metastasis suppressor protein Nm23-H1. J Virol 83(10):4871–4883. CrossRefGoogle Scholar
  31. 31.
    Murakami M, Lan K, Subramanian C, Robertson ES (2005) Epstein-Barr virus nuclear antigen 1 interacts with Nm23-H1 in lymphoblastoid cell lines and inhibits its ability to suppress cell migration. J Virol 79(3):1559–1568. CrossRefGoogle Scholar
  32. 32.
    Qin Z, Dai L, Toole B, Robertson E, Parsons C (2011) Regulation of Nm23-H1 and cell invasiveness by Kaposi’s sarcoma-associated herpesvirus. J Virol 85(7):3596–3606. CrossRefGoogle Scholar
  33. 33.
    Subramanian C, Cotter MA 2nd, Robertson ES (2001) Epstein-Barr virus nuclear protein EBNA-3C interacts with the human metastatic suppressor Nm23-H1: a molecular link to cancer metastasis. Nat Med 7(3):350–355. CrossRefGoogle Scholar
  34. 34.
    Banerjee A, Ray RB, Ray R (2010) Oncogenic potential of hepatitis C virus proteins. Viruses 2(9):2108–2133. CrossRefGoogle Scholar
  35. 35.
    Feng X, Xiu B, Xu L, Yang X, He J, Leong D, He F, Zhang H (2011) Hepatitis C virus core protein promotes the migration and invasion of hepatocyte via activating transcription of extracellular matrix metalloproteinase inducer. Virus Res 158(1):146–153. CrossRefGoogle Scholar
  36. 36.
    Saha A, Robertson ES (2011) Functional modulation of the metastatic suppressor Nm23-H1 by oncogenic viruses. FEBS Lett 585(20):3174–3184. CrossRefGoogle Scholar
  37. 37.
    Fujimoto Y, Ohtake T, Nishimori H, Ikuta K, Ohhira M, Ono M, Kohgo Y (1998) Reduced expression and rare genomic alteration of nm23-H1in human hepatocellular carcinoma and hepatoma cell lines. J Gastroenterol 33(3):368–375. CrossRefGoogle Scholar
  38. 38.
    Nakayama T, Ohtsuru A, Nakao K, Shima M, Nakata K, Watanabe K, Ishii N, Kimura N, Nagataki S (1992) Expression in human hepatocellular carcinoma of nucleoside diphosphate kinase, a homologue of the nm23 gene product. J Natl Cancer Inst 84(17):1349–1354CrossRefGoogle Scholar
  39. 39.
    Gawlik K, Gallay PA (2014) HCV core protein and virus assembly: what we know without structures. Immunol Res 60(1):1–10. CrossRefGoogle Scholar
  40. 40.
    Dumas C, Lascu I, Morera S, Glaser P, Fourme R, Wallet V, Lacombe ML, Veron M, Janin J (1992) X-ray structure of nucleoside diphosphate kinase. EMBO J 11(9):3203–3208CrossRefGoogle Scholar
  41. 41.
    Kim YI, Park S, Jeoung DI, Lee H (2003) Point mutations affecting the oligomeric structure of Nm23-H1 abrogates its inhibitory activity on colonization and invasion of prostate cancer cells. Biochem Biophys Res Commun 307(2):281–289CrossRefGoogle Scholar
  42. 42.
    Fiore LS, Ganguly SS, Sledziona J, Cibull ML, Wang C, Richards DL, Neltner JM, Beach C, McCorkle JR, Kaetzel DM, Plattner R (2014) c-Abl and Arg induce cathepsin-mediated lysosomal degradation of the NM23-H1 metastasis suppressor in invasive cancer. Oncogene 33(36):4508–4520. CrossRefGoogle Scholar
  43. 43.
    Guo WZ, Sugaya S, Satoh M, Tomonaga T, Nomura F, Hiwasa T, Takiguchi M, Kita K, Suzuki N (2009) Nm23-H1 is responsible for SUMO-2-involved DNA synthesis induction after X-ray irradiation in human cells. Arch Biochem Biophys 486(1):81–87. CrossRefGoogle Scholar
  44. 44.
    Bosnar MH, De Gunzburg J, Bago R, Brecevic L, Weber I, Pavelic J (2004) Subcellular localization of A and B Nm23/NDPK subunits. Exp Cell Res 298(1):275–284. CrossRefGoogle Scholar
  45. 45.
    Khera L, Paul C, Kaul R (2018) Hepatitis C Virus mediated metastasis in hepatocellular carcinoma as a therapeutic target for cancer management. Curr Drug Metab. Google Scholar
  46. 46.
    Simmonds P (2004) Genetic diversity and evolution of hepatitis C virus—15 years on. J Gen Virol 85(Pt 11):3173–3188. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of MicrobiologyUniversity of DelhiNew DelhiIndia
  2. 2.Department of Molecular Cell BiologyWeizman Institute of ScienceRehovotIsrael

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