Nuclear/nucleolar localization properties of C-terminal nucleocapsid protein of SARS coronavirus

Khalid A. Timani, Qingjiao Liao, Linbai Ye, Yingchun Zeng, Jing Liu, Yi Zheng, Li Ye, Xiaojun Yang, Kong Lingbao, Jingrong Gao, Ying Zhu

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Abstract

A novel coronavirus (CoV) has recently been identified as the aetiological agent of severe acute respiratory syndrome (SARS). Nucleocapsid (N) proteins of the Coronaviridae family have no discernable homology, but they share a common nucleolar-cytoplasmic distribution pattern. There are three putative nuclear localization signal (NLS) motifs present in the N. To determine the role of these putative NLSs in the intracellular localization of the SARS-CoV N, we performed a confocal microscopy analysis using rabbit anti-N antisera. In this report, we show that the wild type N was distributed mainly in the cytoplasm. The N-terminal of the N, which contains the NLS1 (aa38-44), was localized to the nucleus. The C-terminus of the N, which contains both NLS2 (aa257-265) and NLS3 (aa369-390) was localized to the cytoplasm and the nucleolus. Results derived from analysis of various deletion mutations show that the region containing amino acids 226-289 is able to mediate nucleolar localization. The deletion of two hydrophobic regions that flanked the NLS3 recovered its activity and localized to the nucleus. Furthermore, deletion of leucine rich region (220-LALLLLDRLNRL) resulted in the accumulation of N to the cytoplasm and nucleolus, and when fusing this peptide to EGFP localization was cytoplasmic, suggesting that the N may act as a shuttle protein. Differences in nuclear/nucleolar localization properties of N from other members of coronavirus family suggest a unique function for N, which may play an important role in the pathogenesis of SARS.

Original languageEnglish
Pages (from-to)23-34
Number of pages12
JournalVirus Research
Volume114
Issue number1-2
DOIs
StatePublished - 1 Dec 2005

Fingerprint

Nucleocapsid Proteins
Severe Acute Respiratory Syndrome
Coronavirus
Cytoplasm
Coronaviridae
Nuclear Localization Signals
Sequence Deletion
Confocal Microscopy
Leucine
Immune Sera
Rabbits
Amino Acids
Peptides
Proteins

Keywords

  • Cell cycle arrest
  • Nuclear localization signal
  • Nucleocapsid protein
  • Nucleolar localization
  • SARS-CoV

Cite this

Timani, Khalid A. ; Liao, Qingjiao ; Ye, Linbai ; Zeng, Yingchun ; Liu, Jing ; Zheng, Yi ; Ye, Li ; Yang, Xiaojun ; Lingbao, Kong ; Gao, Jingrong ; Zhu, Ying. / Nuclear/nucleolar localization properties of C-terminal nucleocapsid protein of SARS coronavirus. In: Virus Research. 2005 ; Vol. 114, No. 1-2. pp. 23-34.
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title = "Nuclear/nucleolar localization properties of C-terminal nucleocapsid protein of SARS coronavirus",
abstract = "A novel coronavirus (CoV) has recently been identified as the aetiological agent of severe acute respiratory syndrome (SARS). Nucleocapsid (N) proteins of the Coronaviridae family have no discernable homology, but they share a common nucleolar-cytoplasmic distribution pattern. There are three putative nuclear localization signal (NLS) motifs present in the N. To determine the role of these putative NLSs in the intracellular localization of the SARS-CoV N, we performed a confocal microscopy analysis using rabbit anti-N antisera. In this report, we show that the wild type N was distributed mainly in the cytoplasm. The N-terminal of the N, which contains the NLS1 (aa38-44), was localized to the nucleus. The C-terminus of the N, which contains both NLS2 (aa257-265) and NLS3 (aa369-390) was localized to the cytoplasm and the nucleolus. Results derived from analysis of various deletion mutations show that the region containing amino acids 226-289 is able to mediate nucleolar localization. The deletion of two hydrophobic regions that flanked the NLS3 recovered its activity and localized to the nucleus. Furthermore, deletion of leucine rich region (220-LALLLLDRLNRL) resulted in the accumulation of N to the cytoplasm and nucleolus, and when fusing this peptide to EGFP localization was cytoplasmic, suggesting that the N may act as a shuttle protein. Differences in nuclear/nucleolar localization properties of N from other members of coronavirus family suggest a unique function for N, which may play an important role in the pathogenesis of SARS.",
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Timani, KA, Liao, Q, Ye, L, Zeng, Y, Liu, J, Zheng, Y, Ye, L, Yang, X, Lingbao, K, Gao, J & Zhu, Y 2005, 'Nuclear/nucleolar localization properties of C-terminal nucleocapsid protein of SARS coronavirus', Virus Research, vol. 114, no. 1-2, pp. 23-34. https://doi.org/10.1016/j.virusres.2005.05.007

Nuclear/nucleolar localization properties of C-terminal nucleocapsid protein of SARS coronavirus. / Timani, Khalid A.; Liao, Qingjiao; Ye, Linbai; Zeng, Yingchun; Liu, Jing; Zheng, Yi; Ye, Li; Yang, Xiaojun; Lingbao, Kong; Gao, Jingrong; Zhu, Ying.

In: Virus Research, Vol. 114, No. 1-2, 01.12.2005, p. 23-34.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Nuclear/nucleolar localization properties of C-terminal nucleocapsid protein of SARS coronavirus

AU - Timani, Khalid A.

AU - Liao, Qingjiao

AU - Ye, Linbai

AU - Zeng, Yingchun

AU - Liu, Jing

AU - Zheng, Yi

AU - Ye, Li

AU - Yang, Xiaojun

AU - Lingbao, Kong

AU - Gao, Jingrong

AU - Zhu, Ying

PY - 2005/12/1

Y1 - 2005/12/1

N2 - A novel coronavirus (CoV) has recently been identified as the aetiological agent of severe acute respiratory syndrome (SARS). Nucleocapsid (N) proteins of the Coronaviridae family have no discernable homology, but they share a common nucleolar-cytoplasmic distribution pattern. There are three putative nuclear localization signal (NLS) motifs present in the N. To determine the role of these putative NLSs in the intracellular localization of the SARS-CoV N, we performed a confocal microscopy analysis using rabbit anti-N antisera. In this report, we show that the wild type N was distributed mainly in the cytoplasm. The N-terminal of the N, which contains the NLS1 (aa38-44), was localized to the nucleus. The C-terminus of the N, which contains both NLS2 (aa257-265) and NLS3 (aa369-390) was localized to the cytoplasm and the nucleolus. Results derived from analysis of various deletion mutations show that the region containing amino acids 226-289 is able to mediate nucleolar localization. The deletion of two hydrophobic regions that flanked the NLS3 recovered its activity and localized to the nucleus. Furthermore, deletion of leucine rich region (220-LALLLLDRLNRL) resulted in the accumulation of N to the cytoplasm and nucleolus, and when fusing this peptide to EGFP localization was cytoplasmic, suggesting that the N may act as a shuttle protein. Differences in nuclear/nucleolar localization properties of N from other members of coronavirus family suggest a unique function for N, which may play an important role in the pathogenesis of SARS.

AB - A novel coronavirus (CoV) has recently been identified as the aetiological agent of severe acute respiratory syndrome (SARS). Nucleocapsid (N) proteins of the Coronaviridae family have no discernable homology, but they share a common nucleolar-cytoplasmic distribution pattern. There are three putative nuclear localization signal (NLS) motifs present in the N. To determine the role of these putative NLSs in the intracellular localization of the SARS-CoV N, we performed a confocal microscopy analysis using rabbit anti-N antisera. In this report, we show that the wild type N was distributed mainly in the cytoplasm. The N-terminal of the N, which contains the NLS1 (aa38-44), was localized to the nucleus. The C-terminus of the N, which contains both NLS2 (aa257-265) and NLS3 (aa369-390) was localized to the cytoplasm and the nucleolus. Results derived from analysis of various deletion mutations show that the region containing amino acids 226-289 is able to mediate nucleolar localization. The deletion of two hydrophobic regions that flanked the NLS3 recovered its activity and localized to the nucleus. Furthermore, deletion of leucine rich region (220-LALLLLDRLNRL) resulted in the accumulation of N to the cytoplasm and nucleolus, and when fusing this peptide to EGFP localization was cytoplasmic, suggesting that the N may act as a shuttle protein. Differences in nuclear/nucleolar localization properties of N from other members of coronavirus family suggest a unique function for N, which may play an important role in the pathogenesis of SARS.

KW - Cell cycle arrest

KW - Nuclear localization signal

KW - Nucleocapsid protein

KW - Nucleolar localization

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