Archives of Virology

, Volume 163, Issue 6, pp 1531–1547 | Cite as

New tetrameric forms of the rotavirus NSP4 with antiparallel helices

  • Sushant Kumar
  • Raghavendra Ramappa
  • Kiranmayee Pamidimukkala
  • C. D. Rao
  • K. Suguna
Original Article


Rotavirus nonstructural protein 4, the first viral enterotoxin to be identified, is a multidomain, multifunctional glycoprotein. Earlier, we reported a Ca2+-bound coiled-coil tetrameric structure of the diarrhea-inducing region of NSP4 from the rotavirus strains SA11 and I321 and a Ca2+-free pentameric structure from the rotavirus strain ST3, all with a parallel arrangement of α-helices. pH was found to determine the oligomeric state: a basic pH favoured a tetramer, whereas an acidic pH favoured a pentamer. Here, we report two novel forms of the coiled-coil region of NSP4 from the bovine rotavirus strains MF66 and NCDV. These crystallized at acidic pH, forming antiparallel coiled-coil tetrameric structures without any bound Ca2+ ion. Structural and mutational studies of the coiled-coil regions of NSP4 revealed that the nature of the residue at position 131 (Tyr/His) plays an important role in the observed structural diversity.



This research was funded by the Department of Biotechnology (DBT) – Indian Institute of Science (IISc) Partnership Program for Advanced Research in Biological Sciences and Bioengineering. Diffraction data were collected at the X-ray Facility for Structural Biology at the Molecular Biophysics Unit, IISc, supported by the Department of Science and Technology, Government of India. SK acknowledges the Council of Scientific and Industrial Research, Government of India, for providing a research fellowship.

Compliance of ethical standards

Conflict of interest

The authors declare no conflict of interest.

Ethical approval

Necessary approvals from Institutional Biosafety Committee and Animal Ethics Committee were obtained prior to the start of animal experiments. This article does not contain any studies with human participants performed by any of the authors.

Supplementary material

705_2018_3753_MOESM1_ESM.pdf (28 kb)
Supplementary Material 1 Lack of correlation between the diarrhea-inducing ability of NSP4ΔN72 deletion mutant proteins and the virulence of rotavirus strains. The DD50 of ΔN72 recombinant protein from the bovine strain MF66 was determined in this study. The values for the protein from other strains were reported by us previously (PDF 27 kb)
705_2018_3753_MOESM2_ESM.pdf (895 kb)
Supplementary Material 2 Sequence alignment of NSP4 from different strains of rotavirus. Residues at positions 120, 131 and 133 are highlighted in yellow, green and blue, respectively (PDF 894 kb)


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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  • Sushant Kumar
    • 1
  • Raghavendra Ramappa
    • 2
  • Kiranmayee Pamidimukkala
    • 2
  • C. D. Rao
    • 2
  • K. Suguna
    • 1
  1. 1.Molecular Biophysics UnitIndian Institute of ScienceBangaloreIndia
  2. 2.Department of Microbiology and Cell BiologyIndian Institute of ScienceBangaloreIndia

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