Archives of Virology

, Volume 157, Issue 4, pp 635–645 | Cite as

SARS-CoV nucleocapsid protein interacts with cellular pyruvate kinase protein and inhibits its activity

  • Wei-Yen Wei
  • Hui-Chun Li
  • Chiung-Yao Chen
  • Chee-Hing Yang
  • Shen-Kao Lee
  • Chia-Wen Wang
  • Hsin-Chieh Ma
  • Yue-Li Juang
  • Shih-Yen Lo
Original Article
First Online:
Received:
Accepted:

Abstract

The pathogenesis of SARS-CoV remains largely unknown. To study the function of the SARS-CoV nucleocapsid protein, we have conducted a yeast two-hybrid screening experiment to identify cellular proteins that may interact with the SARS-CoV nucleocapsid protein. Pyruvate kinase (liver) was found to interact with SARS-CoV nucleocapsid protein in this experiment. The binding domains of these two proteins were also determined using the yeast two-hybrid system. The physical interaction between the SARS-CoV nucleocapsid and cellular pyruvate kinase (liver) proteins was further confirmed by GST pull-down assay, co-immunoprecipitation assay and confocal microscopy. Cellular pyruvate kinase activity in hepatoma cells was repressed by SARS-CoV nucleocapsid protein in either transiently transfected or stably transfected cells. PK deficiency in red blood cells is known to result in human hereditary non-spherocytic hemolytic anemia. It is reasonable to assume that an inhibition of PKL activity due to interaction with SARS-CoV N protein is likely to cause the death of the hepatocytes, which results in the elevation of serum alanine aminotransferase and liver dysfunction noted in most SARS patients. Thus, our results suggest that SARS-CoV could reduce pyruvate kinase activity via its nucleocapsid protein, and this may in turn cause disease.

Keywords

Pyruvate Kinase HuH7 Cell Severe Acute Respiratory Syndrome Pyruvate Kinase Activity Murine Hepatitis Virus Type 
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.
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Notes

Acknowledgments

We thank Dr. Li-Kuang Chen for providing SARS-CoV cDNA. RNAi reagents were obtained from the National RNAi Core Facility located at the Institute of Molecular Biology/Genomic Research Center, Academia Sinica, supported by grants from the NSC National Research Program for Genomic Medicine (NSC 94-3112-B-001-003 and NSC 94-3112-B-001-018-Y). This work was supported by grants from the National Science Council of Taiwan (NSC 98-2320-B-320-001-MY3) and from the Tzu Chi University (TCIRP96004-05) to Dr. Shih-Yen Lo.

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Wei-Yen Wei
    • 1
  • Hui-Chun Li
    • 1
    • 2
  • Chiung-Yao Chen
    • 1
  • Chee-Hing Yang
    • 3
  • Shen-Kao Lee
    • 3
  • Chia-Wen Wang
    • 3
  • Hsin-Chieh Ma
    • 2
  • Yue-Li Juang
    • 1
    • 2
  • Shih-Yen Lo
    • 1
    • 2
    • 3
    • 4
  1. 1.Graduate Institute of Molecular and Cellular BiologyTzu Chi UniversityHualienTaiwan
  2. 2.Graduate Institute of Medical SciencesTzu Chi UniversityHualienTaiwan
  3. 3.Graduate Institute of Medical BiotechnologyTzu Chi UniversityHualienTaiwan
  4. 4.Department of Laboratory MedicineBuddhist Tzu Chi General HospitalHualienTaiwan

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