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Subcellular localization of the severe acute respiratory syndrome coronavirus nucleocapsid protein

Paul Heinen^4,

¹ Institute of Molecular and Cellular Biology, Faculty of Biological Sciences, Garstang Building, University of Leeds, Leeds LS2 9JT, UK
² Department of Molecular and Cell Biology, Centro Nacional de Biotecnología (CNB, CSIC), Campus Univ. Autonoma, 3 Darwin Street, Cantoblanco, 28049 Madrid, Spain
³ Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK
⁴ Health Protection Agency, London NW9 5HT, UK

Correspondence
Julian A. Hiscox
j.a.hiscox{at}leeds.ac.uk

The coronavirus nucleocapsid (N) protein is a viral RNA-binding protein with multiple functions in terms of virus replication and modulating cell signalling pathways. N protein is composed of three distinct regions containing RNA-binding motif(s), and appropriate signals for modulating cell signalling. The subcellular localization of severe acute respiratory syndrome coronavirus (SARS-CoV) N protein was studied. In infected cells, SARS-CoV N protein localized exclusively to the cytoplasm. In contrast to the avian coronavirus N protein, overexpressed SARS-CoV N protein remained principally localized to the cytoplasm, with very few cells exhibiting nucleolar localization. Bioinformatic analysis and deletion mutagenesis coupled to confocal microscopy and live-cell imaging, revealed that SARS-CoV N protein regions I and III contained nuclear localization signals and region II contained a nucleolar retention signal. However, cytoplasmic localization was directed by region III and was the dominant localization signal in the protein.

Present address: Institute for Animal Health, Pirbright GU24 0NF, UK.

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