Archives of Virology

, Volume 164, Issue 4, pp 1173–1180 | Cite as

The coordinating role of the human norovirus minor capsid protein VP2 is essential to functional change and nuclear localization of the major capsid protein VP1

  • Zhili Liu
  • Min Zhang
  • Zhen Shen
  • Huifen Chen
  • Wanju Zhang
  • Xiaoqing Xu
  • Zelin Lai
  • Wenqin Sun
  • Zheng ZhaoEmail author
  • Jun ZhangEmail author
Original Article


Global outbreaks of norovirus (NOV) gastroenteritis are associated with the most prevalent genotype, GII.4. Mutations in the protruding domain 2 (P2 domain) of the norovirus major capsid protein (VP1) result in the emergence of various NOV variants, however, it is unclear whether the minor capsid protein (VP2) also affects the generation of VP1 variants. In this study, using a human 293T expression system, we investigated the interactions of VP1 and VP2 of three GII.4 strains, focusing on the changes in expression and cellular localization. We found that co-transfection with VP1 and VP2 leads to a significant increase in expression of both proteins compared to that in cells transfected with VP1 or VP2 alone. In contrast to VP1 expressed in the absence of VP2, which was dispersed throughout the cytosol, VP2 expressed in the absence of VP1 was found to be located in the nucleus. This could be attributed to a predicted specific nuclear localization signal found in this gene. When both proteins were expressed, VP1 was found together with VP2 in the nucleus. These results thus suggest that the VP2 of GII.4 NOVs affects the function and cellular location of VP1 and that, with the cooperation of VP2, VP1 could play a critical role in affecting cell functions by impairing the downstream transcriptional signaling and chromatin remodeling in the cell nuclei.



This study was funded by the National Natural Science Foundation of China (81470829 to JZ).

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_4192_MOESM1_ESM.xlsx (18 kb)
Supplementary material 1 (XLSX 17 kb)


  1. 1.
    Ahmed SM, Hall AJ, Robinson AE, Verhoef L, Premkumar P, Parashar UD, Koopmans M, Lopman BA (2014) Global prevalence of norovirus in cases of gastroenteritis: a systematic review and meta-analysis. Lancet Infect Dis 14:725–730CrossRefGoogle Scholar
  2. 2.
    Ba ANN, Pogoutse A, Provart N, Moses MA (2009) NLStradamus: a simple Hidden Markov Model for nuclear localization signal prediction. BMC Bioinformatics 10:202CrossRefGoogle Scholar
  3. 3.
    Bertolotticiarlet A, White LJ, Chen R, Prasad BVV, Estes MK (2002) Structural Requirements for the Assembly of Norwalk Virus-Like Particles. J Virol 76:4044CrossRefGoogle Scholar
  4. 4.
    Bertolotticiarlet A, Crawford SE, Hutson AM, Estes MK (2003) The 3′ End of Norwalk Virus mRNA Contains Determinants That Regulate the Expression and Stability of the Viral Capsid Protein VP1: a Novel Function for the VP2 Protein. J Virol 77:11603–11615CrossRefGoogle Scholar
  5. 5.
    Chan CW, Lee N, Ho WS, Law OK, Lau CK, Tsui KW, Sung JY (2012) Covariation of Major and Minor Viral Capsid Proteins in Norovirus Genogroup II Genotype 4 Strains. J Virol 86:1227–1232CrossRefGoogle Scholar
  6. 6.
    Chanit W, Thongprachum A, Okitsu S, Mizuguchi M, Ushijima H (2013) Genetic analysis and homology modeling of capsid protein of norovirus GII.14. J Med Virol 86:329–334CrossRefGoogle Scholar
  7. 7.
    Crooks GE, Hon G, Chandonia J-M, Brenner SE (2004) WebLogo: a sequence logo generator. Genome Res 14:1188–1190CrossRefGoogle Scholar
  8. 8.
    Donaldson EF, Lindesmith LC, Lobue AD, Baric RS (2010) Viral shape-shifting: norovirus evasion of the human immune system. Nat Rev Microbiol 8:231–241CrossRefGoogle Scholar
  9. 9.
    Franz O, Manfred O (2005) Identification of an unconventional nuclear localization signal in human ribosomal protein S2. Biochem Biophys Res Commun 335:146–153CrossRefGoogle Scholar
  10. 10.
    Glass PJ, White LJ, Ball JM, Leparcgoffart I, Hardy ME, Estes MK (2000) Norwalk virus open reading frame 3 encodes a minor structural protein. J Virol 74:6581CrossRefGoogle Scholar
  11. 11.
    Graaf MD, Beek JV, Koopmans MPG (2016) Human norovirus transmission and evolution in a changing world. Nat Rev Microbiol 14:421CrossRefGoogle Scholar
  12. 12.
    Hoa Tran TN, Trainor E, Nakagomi T, Cunliffe NA, Nakagomi O (2013) Molecular epidemiology of noroviruses associated with acute sporadic gastroenteritis in children: global distribution of genogroups, genotypes and GII.4 variants. J Clin Virol 56:185–193CrossRefGoogle Scholar
  13. 13.
    Jiang X, Wang M, Graham DY, Estes MK (1992) Expression, self-assembly, and antigenicity of the Norwalk virus capsid protein. J Virol 66:6527–6532Google Scholar
  14. 14.
    Jiang X, Wang M, Wang K, Estes MK (1993) Sequence and Genomic Organization of Norwalk Virus. Virology 195:51–61CrossRefGoogle Scholar
  15. 15.
    Kapikian AZ, Wyatt RG, Dolin R, Thornhill TS, Kalica AR, Chanock RM (1972) Visualization by immune electron microscopy of a 27-nm particle associated with acute infectious nonbacterial gastroenteritis. J Virol 10:1075–1081Google Scholar
  16. 16.
    Kari D, Lindesmith LC, Donaldson EF, Veronica C, Martina B, Davide C, Jesica S, Antonio L, Jan V, Baric RS (2013) Emergence of New Pandemic GII.4 Sydney Norovirus Strain Correlates With Escape From Herd Immunity. J Infect Dis 208:1877–1887CrossRefGoogle Scholar
  17. 17.
    Kretsovali A, Spilianakis C, Dimakopoulos A, Makatounakis T, Papamatheakis J (2001) Self-association of class II transactivator correlates with its intracellular localization and transactivation. J Biol Chem 276:32191–32197CrossRefGoogle Scholar
  18. 18.
    Lin Y, Fengling L, Lianzhu W, Yuxiu Z, Yanhua J (2014) Function of VP2 protein in the stability of the secondary structure of virus-like particles of genogroup II norovirus at different pH levels: Function of VP2 protein in the stability of NoV VLPs. J Microbiol 52:970CrossRefGoogle Scholar
  19. 19.
    Lindesmith LC, Verónica C, Jesica S, Kari D, Donaldson EF, Jan V, Baric RS (2013) Emergence of a norovirus GII.4 strain correlates with changes in evolving blockade epitopes. J Virol 87:2803–2813CrossRefGoogle Scholar
  20. 20.
    Lindsay L, Wolter J, Coster ID, Damme PV, Verstraeten T (2015) A decade of norovirus disease risk among older adults in upper-middle and high income countries: a systematic review. BMC Infect Dis 15:425CrossRefGoogle Scholar
  21. 21.
    Lischka P, Sorg G, Kann M, Winkler M, Stamminger T (2003) A nonconventional nuclear localization signal within the UL84 protein of human cytomegalovirus mediates nuclear import via the importin alpha/beta pathway. J Virol 77:3734CrossRefGoogle Scholar
  22. 22.
    Motoya T, Nagasawa K, Matsushima Y, Nagata N, Ryo A, Sekizuka T, Yamashita A, Kuroda M, Morita Y, Suzuki Y (2017) Molecular evolution of the VP1 gene in human norovirus GII.4 variants in 1974–2015. Front Microbiol 8:2399CrossRefGoogle Scholar
  23. 23.
    Patel MM, Hall AJ, Vinjé J, Parashar UD (2009) Noroviruses: a comprehensive review. J Clin Virol 44:1–8CrossRefGoogle Scholar
  24. 24.
    Sabrià A, Pintó RM, Bosch A, Bartolomé R, Cornejo T, Torner N, Martínez A, De SM, Domínguez A, Guix S (2014) Molecular and clinical epidemiology of norovirus outbreaks in Spain during the emergence of GII.4 2012 variant. J Clin Virol 60:96–104CrossRefGoogle Scholar
  25. 25.
    Sosnovtsev SV, Green KY (2000) Identification and genomic mapping of the ORF3 and VPg proteins in feline calicivirus virions. Virology 277:193–203CrossRefGoogle Scholar
  26. 26.
    Sosnovtsev SV, Belliot G, Chang KO, Onwudiwe O, Green KY (2005) Feline calicivirus VP2 is essential for the production of infectious virions. J Virol 79:4012–4024CrossRefGoogle Scholar
  27. 27.
    Vinjé J (2015) Advances in laboratory methods for detection and typing of norovirus. J Clin Microbiol 53:373–381CrossRefGoogle Scholar
  28. 28.
    Vongpunsawad S, Prasad BVV, Estes MK (2013) Norwalk Virus Minor Capsid Protein VP2 Associates within the VP1 Shell Domain. J Virol 87:4818CrossRefGoogle Scholar
  29. 29.
    Zhang J, Shen Z, Zhu Z, Zhang W, Chen H, Qian F, Chen H, Wang G, Wang M, Hu Y (2015) Genotype distribution of norovirus around the emergence of Sydney_2012 and the antigenic drift of contemporary GII.4 epidemic strains. J Clin Virol 72:95–101CrossRefGoogle Scholar
  30. 30.
    Zhu J, Qiu Z, Wiese C, Ishii Y, Friedrichsen J, Rajashekara G, Splitter GA (2005) Nuclear and mitochondrial localization signals overlap within bovine herpesvirus 1 tegument protein VP22. J Biol Chem 280:16038CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Key Laboratory of Brain Functional Genomics (East China Normal University), Ministry of Education School of Life SciencesEast China Normal UniversityShanghaiChina
  2. 2.Department of Clinical LaboratoryShanghai Public Health Clinical Center Affiliated to Fudan UniversityShanghaiChina
  3. 3.Department of Laboratory Medicine, Renji Hospital ShanghaiJiaotong University School of MedicineShanghaiChina
  4. 4.Department of Clinical Laboratory, Shanghai First Maternity and Infant HospitalTongji University School of MedicineShanghaiChina
  5. 5.Department of Pathogen Diagnosis and BiosafetyShanghai Public Health Clinical Center Affiliated to Fudan UniversityShanghaiChina

Personalised recommendations