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Archives of Virology

, Volume 156, Issue 12, pp 2163–2172 | Cite as

Investigations on the RNA binding and phosphorylation of groundnut bud necrosis virus nucleocapsid protein

  • Amruta S. Bhat
  • H. S. SavithriEmail author
Original Article

Abstract

Groundnut bud necrosis virus belongs to the genus Tospovirus, infects a wide range of crop plants and causes severe losses. To understand the role of the nucleocapsid protein in the viral life cycle, the protein was overexpressed in E. coli and purified by Ni-NTA chromatography. The purified N protein was well folded and was predominantly alpha-helical. Deletion analysis revealed that the C-terminal unfolded region of the N protein was involved in RNA binding. Furthermore, the N protein could be phosphorylated in vitro by Nicotiana benthamiana plant sap and by purified recombinant kinases such as protein kinase CK2 and calcium-dependent protein kinase. This is the first report of phoshphorylation of a nucleocapsid protein in the family Bunyaviridae. The possible implications of the present findings for the viral life cycle are discussed.

Keywords

Infectious Bronchitis Virus Tomato Spotted Wilt Virus Nucleocapsid Protein Viral Life Cycle Rift Valley Fever 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations

GBNV

Groundnut bud necrosis virus

TSWV

Tomato spotted wilt virus

RNP

Ribonucleoprotein

N protein

Nucleocapsid protein

CK2

Protein kinase CK2

CDPK

Calcium-dependent protein kinase

Notes

Acknowledgments

We thank the Department of Biotechnology (DBT) and Department of Science and Technology, New Delhi, India for financial support. ASB acknowledges DBT for a postdoctoral fellowship. We are grateful to Prof. Kristiina Makinen, University of Helsinki, for providing the tobacco CK2 clone. We thank Mr. Ajay and Prof. C. Jayabaskaran, IISc, for a gift of purified recombinant chickpea CDPK.

Supplementary material

705_2011_1110_MOESM1_ESM.pdf (26 kb)
Supplementary material 1 (PDF 25 kb)
705_2011_1110_MOESM2_ESM.pdf (43 kb)
Supplementary material 2 (PDF 43 kb)
705_2011_1110_MOESM3_ESM.pdf (54 kb)
Supplementary material 3 (PDF 53 kb)

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.Department of BiochemistryIndian Institute of ScienceBangaloreIndia

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