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1 Institute of Virology and Immunology, University of Würzburg, Versbacher Str. 7, 97078 Würzburg, Germany
2 Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, Leiden, The Netherlands
3 Institute for Medical Virology, Johann Wolfgang Goethe University, Frankfurt (Main), Germany
Correspondence
John Ziebuhr
j.ziebuhr{at}mail.uni-wuerzburg.de
A novel coronavirus is the causative agent of the current epidemic of severe acute respiratory syndrome (SARS). Coronaviruses are exceptionally large RNA viruses and employ complex regulatory mechanisms to express their genomes. Here, we determined the sequence of SARS coronavirus (SARS-CoV), isolate Frankfurt 1, and characterized key RNA elements and protein functions involved in viral genome expression. Important regulatory mechanisms, such as the (discontinuous) synthesis of eight subgenomic mRNAs, ribosomal frameshifting and post-translational proteolytic processing, were addressed. Activities of three SARS coronavirus enzymes, the helicase and two cysteine proteinases, which are known to be critically involved in replication, transcription and/or post-translational polyprotein processing, were characterized. The availability of recombinant forms of key replicative enzymes of SARS coronavirus should pave the way for high-throughput screening approaches to identify candidate inhibitors in compound libraries.
Present address: Research Department, Cantonal Hospital, St Gallen, Switzerland.
Published ahead of print on 19 June 2003 as DOI 10.1099/vir.0.19424-0.
The nucleotide sequence of SARS-CoV, isolate Frankfurt 1, has been deposited in GenBank, accession no. AY291315.
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J. R. St-Jean, H. Jacomy, M. Desforges, A. Vabret, F. Freymuth, and P. J. Talbot Human Respiratory Coronavirus OC43: Genetic Stability and Neuroinvasion J. Virol., August 15, 2004; 78(16): 8824 - 8834. [Abstract] [Full Text] [PDF] |
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A. E. Gorbalenya, E. J. Snijder, and W. J. M. Spaan Severe Acute Respiratory Syndrome Coronavirus Phylogeny: toward Consensus J. Virol., August 1, 2004; 78(15): 7863 - 7866. [Full Text] [PDF] |
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B. C. Fielding, Y.-J. Tan, S. Shuo, T. H. P. Tan, E.-E. Ooi, S. G. Lim, W. Hong, and P.-Y. Goh Characterization of a Unique Group-Specific Protein (U122) of the Severe Acute Respiratory Syndrome Coronavirus J. Virol., July 15, 2004; 78(14): 7311 - 7318. [Abstract] [Full Text] [PDF] |
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K. A. Ivanov and J. Ziebuhr Human Coronavirus 229E Nonstructural Protein 13: Characterization of Duplex-Unwinding, Nucleoside Triphosphatase, and RNA 5'-Triphosphatase Activities J. Virol., July 15, 2004; 78(14): 7833 - 7838. [Abstract] [Full Text] [PDF] |
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L. Gillim-Ross, J. Taylor, D. R. Scholl, J. Ridenour, P. S. Masters, and D. E. Wentworth Discovery of Novel Human and Animal Cells Infected by the Severe Acute Respiratory Syndrome Coronavirus by Replication-Specific Multiplex Reverse Transcription-PCR J. Clin. Microbiol., July 1, 2004; 42(7): 3196 - 3206. [Abstract] [Full Text] [PDF] |
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Y.-J. Tan, E. Teng, S. Shen, T. H. P. Tan, P.-Y. Goh, B. C. Fielding, E.-E. Ooi, H.-C. Tan, S. G. Lim, and W. Hong A Novel Severe Acute Respiratory Syndrome Coronavirus Protein, U274, Is Transported to the Cell Surface and Undergoes Endocytosis J. Virol., July 1, 2004; 78(13): 6723 - 6734. [Abstract] [Full Text] [PDF] |
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H. Zhang, G. Wang, J. Li, Y. Nie, X. Shi, G. Lian, W. Wang, X. Yin, Y. Zhao, X. Qu, et al. Identification of an Antigenic Determinant on the S2 Domain of the Severe Acute Respiratory Syndrome Coronavirus Spike Glycoprotein Capable of Inducing Neutralizing Antibodies J. Virol., July 1, 2004; 78(13): 6938 - 6945. [Abstract] [Full Text] [PDF] |
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J. Shi, Z. Wei, and J. Song Dissection Study on the Severe Acute Respiratory Syndrome 3C-like Protease Reveals the Critical Role of the Extra Domain in Dimerization of the Enzyme: DEFINING THE EXTRA DOMAIN AS A NEW TARGET FOR DESIGN OF HIGHLY SPECIFIC PROTEASE INHIBITORS J. Biol. Chem., June 4, 2004; 279(23): 24765 - 24773. [Abstract] [Full Text] [PDF] |
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T. Hertzig, E. Scandella, B. Schelle, J. Ziebuhr, S. G. Siddell, B. Ludewig, and V. Thiel Rapid identification of coronavirus replicase inhibitors using a selectable replicon RNA J. Gen. Virol., June 1, 2004; 85(6): 1717 - 1725. [Abstract] [Full Text] [PDF] |
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K. A. Ivanov, V. Thiel, J. C. Dobbe, Y. van der Meer, E. J. Snijder, and J. Ziebuhr Multiple Enzymatic Activities Associated with Severe Acute Respiratory Syndrome Coronavirus Helicase J. Virol., June 1, 2004; 78(11): 5619 - 5632. [Abstract] [Full Text] [PDF] |
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M. R. Denison, B. Yount, S. M. Brockway, R. L. Graham, A. C. Sims, X. Lu, and R. S. Baric Cleavage between Replicase Proteins p28 and p65 of Mouse Hepatitis Virus Is Not Required for Virus Replication J. Virol., June 1, 2004; 78(11): 5957 - 5965. [Abstract] [Full Text] [PDF] |
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A. D. L. Sihoe, R. H. L. Wong, A. T. H. Lee, L. S. Lau, N. Y. Y. Leung, K. I. Law, and A. P. C. Yim Severe Acute Respiratory Syndrome Complicated by Spontaneous Pneumothorax Chest, June 1, 2004; 125(6): 2345 - 2351. [Abstract] [Full Text] [PDF] |
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W.-Y. Choy, S.-G. Lin, P. K.-S. Chan, J. S.-L. Tam, Y.M. D. Lo, I. M.-T. Chu, S.-N. Tsai, M.-Q. Zhong, K.-P. Fung, M. M.-Y. Waye, et al. Synthetic Peptide Studies on the Severe Acute Respiratory Syndrome (SARS) Coronavirus Spike Glycoprotein: Perspective for SARS Vaccine Development Clin. Chem., June 1, 2004; 50(6): 1036 - 1042. [Abstract] [Full Text] [PDF] |
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M.-P. Egloff, F. Ferron, V. Campanacci, S. Longhi, C. Rancurel, H. Dutartre, E. J. Snijder, A. E. Gorbalenya, C. Cambillau, and B. Canard The severe acute respiratory syndrome-coronavirus replicative protein nsp9 is a single-stranded RNA-binding subunit unique in the RNA virus world PNAS, March 16, 2004; 101(11): 3792 - 3796. [Abstract] [Full Text] [PDF] |
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A. M. Bressler and F. S. Nolte Preclinical Evaluation of Two Real-Time, Reverse Transcription-PCR Assays for Detection of the Severe Acute Respiratory Syndrome Coronavirus J. Clin. Microbiol., March 1, 2004; 42(3): 987 - 991. [Abstract] [Full Text] [PDF] |
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E. VAN DEN BORN, A. P. GULTYAEV, and E. J. SNIJDER Secondary structure and function of the 5'-proximal region of the equine arteritis virus RNA genome RNA, March 1, 2004; 10(3): 424 - 437. [Abstract] [Full Text] [PDF] |
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Y.-J. Tan, P.-Y. Goh, B. C. Fielding, S. Shen, C.-F. Chou, J.-L. Fu, H. N. Leong, Y. S. Leo, E. E. Ooi, A. E. Ling, et al. Profiles of Antibody Responses against Severe Acute Respiratory Syndrome Coronavirus Recombinant Proteins and Their Potential Use as Diagnostic Markers Clin. Vaccine Immunol., March 1, 2004; 11(2): 362 - 371. [Abstract] [Full Text] [PDF] |
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X. Xu, Y. Liu, S. Weiss, E. Arnold, S. G. Sarafianos, and J. Ding Molecular model of SARS coronavirus polymerase: implications for biochemical functions and drug design Nucleic Acids Res., December 15, 2003; 31(24): 7117 - 7130. [Abstract] [Full Text] [PDF] |
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J. A. Tanner, R. M. Watt, Y.-B. Chai, L.-Y. Lu, M. C. Lin, J. S. M. Peiris, L. L. M. Poon, H.-F. Kung, and J.-D. Huang The Severe Acute Respiratory Syndrome (SARS) Coronavirus NTPase/Helicase Belongs to a Distinct Class of 5' to 3' Viral Helicases J. Biol. Chem., October 10, 2003; 278(41): 39578 - 39582. [Abstract] [Full Text] [PDF] |
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