This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Li, Y.-I.
Right arrow Articles by Meng, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Li, Y.-I.
Right arrow Articles by Meng, M.

 Previous Article  |  Next Article 

Journal of Virology, December 1998, p. 10093-10099, Vol. 72, No. 12
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Identification and Characterization of the Escherichia coli-Expressed RNA-Dependent RNA Polymerase of Bamboo Mosaic Virus

Yi-Ija Li,1 Yueh-Mei Cheng,2 Yih-Leh Huang,1 Ching-Hsiu Tsai,3 Yau-Heiu Hsu,3 and Menghsiao Meng1,*

Graduate Institute of Agricultural Biotechnology,1 Graduate Institute of Biochemistry,2 and Agricultural Biotechnology Laboratories,3 National Chung Hsing University, Taichung, Taiwan 40227, Republic of China

Received 11 May 1998/Accepted 25 August 1998

Bamboo mosaic virus (BaMV), a member of the potexvirus group, infects primarily members of the Bambusoideae. The open reading frame 1 (ORF1) of BaMV encodes a 155-kDa polypeptide that was postulated to be involved in the replication and the formation of cap structure at the 5' end of the viral genome. To characterize the activities associated with the 155-kDa viral protein, it was expressed in Escherichia coli BL21(DE3) cells with thioredoxin, hexahistidine, and S · Tag fused consecutively at its amino terminus, and the fusion protein was purified by metal affinity chromatography. Several RNA fragments, prepared by in vitro transcription, were tested as substrates for the RNA-dependent RNA polymerase (RdRp) activity. Among them, the expressed fusion enzyme was able to generate a 32P-labeled RNA product when 3'-end RNA fragments of the positive strand or negative strand of BaMV were included in the assay mixture. Dot hybridization assay revealed that the reaction products are complementary to their RNA substrates. Taken together, the evidence suggests that the 155-kDa protein encoded by ORF1 of BaMV has an RdRp activity and should be involved in the replication of BaMV. Mutational analyses demonstrate the importance of the GDD motif in the polymerase activity, and deletion studies suggest that the polymerase activity resides in the carboxyl terminus of the 155-kDa viral protein.

* Corresponding author. Mailing address: Graduate Institute of Agricultural Biotechnology, National Chung Hsing University, 250 Kuo-Kuang Rd., Taichung, Taiwan 40227, Republic of China. Phone: 886-04-2840328. Fax: 886-4-2853527. E-mail: mhmeng{at}dragon.nchu.edu.tw.

Journal of Virology, December 1998, p. 10093-10099, Vol. 72, No. 12
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Draghici, H.-K., Varrelmann, M. (2009). Evidence that the Linker between the Methyltransferase and Helicase Domains of Potato Virus X Replicase Is Involved in Homologous RNA Recombination. J. Virol. 83: 7761-7769 [Abstract] [Full Text]  
  • Cheng, C.-W., Hsiao, Y.-Y., Wu, H.-C., Chuang, C.-M., Chen, J.-S., Tsai, C.-H., Hsu, Y.-H., Wu, Y.-C., Lee, C.-C., Meng, M. (2009). Suppression of Bamboo Mosaic Virus Accumulation by a Putative Methyltransferase in Nicotiana benthamiana. J. Virol. 83: 5796-5805 [Abstract] [Full Text]  
  • Lin, J.-W., Ding, M.-P., Hsu, Y.-H., Tsai, C.-H. (2007). Chloroplast phosphoglycerate kinase, a gluconeogenetic enzyme, is required for efficient accumulation of Bamboo mosaic virus. Nucleic Acids Res 35: 424-432 [Abstract] [Full Text]  
  • Chen, I-H., Chou, W.-J., Lee, P.-Y., Hsu, Y.-H., Tsai, C.-H. (2005). The AAUAAA Motif of Bamboo Mosaic Virus RNA Is Involved in Minus-Strand RNA Synthesis and Plus-Strand RNA Polyadenylation. J. Virol. 79: 14555-14561 [Abstract] [Full Text]  
  • Lin, J.-W., Chiu, H.-N., Chen, I-H., Chen, T.-C., Hsu, Y.-H., Tsai, C.-H. (2005). Structural and Functional Analysis of the cis-Acting Elements Required for Plus-Strand RNA Synthesis of Bamboo Mosaic Virus. J. Virol. 79: 9046-9053 [Abstract] [Full Text]  
  • Huang, Y.-L., Hsu, Y.-H., Han, Y.-T., Meng, M. (2005). mRNA Guanylation Catalyzed by the S-Adenosylmethionine-dependent Guanylyltransferase of Bamboo Mosaic Virus. J. Biol. Chem. 280: 13153-13162 [Abstract] [Full Text]  
  • Wang, H.-H., Yu, H.-H., Wong, S.-M. (2004). Mutation of Phe50 to Ser50 in the 126/183-kDa proteins of Odontoglossum ringspot virus abolishes virus replication but can be complemented and restored by exact reversion. J. Gen. Virol. 85: 2447-2457 [Abstract] [Full Text]  
  • Huang, Y.-L., Han, Y.-T., Chang, Y.-T., Hsu, Y.-H., Meng, M. (2004). Critical Residues for GTP Methylation and Formation of the Covalent m7GMP-Enzyme Intermediate in the Capping Enzyme Domain of Bamboo Mosaic Virus. J. Virol. 78: 1271-1280 [Abstract] [Full Text]  
  • Cheng, J.-H., Peng, C.-W., Hsu, Y.-H., Tsai, C.-H. (2002). The Synthesis of Minus-Strand RNA of Bamboo Mosaic Potexvirus Initiates from Multiple Sites within the Poly(A) Tail. J. Virol. 76: 6114-6120 [Abstract] [Full Text]  
  • Rajendran, K. S., Pogany, J., Nagy, P. D (2002). Comparison of Turnip Crinkle Virus RNA-Dependent RNA Polymerase Preparations Expressed in Escherichia coli or Derived from Infected Plants. J. Virol. 76: 1707-1717 [Abstract] [Full Text]  
  • Li, Y.-I., Shih, T.-W., Hsu, Y.-H., Han, Y.-T., Huang, Y.-L., Meng, M. (2001). The Helicase-Like Domain of Plant Potexvirus Replicase Participates in Formation of RNA 5' Cap Structure by Exhibiting RNA 5'-Triphosphatase Activity. J. Virol. 75: 12114-12120 [Abstract] [Full Text]  
  • Vlot, A. C., Neeleman, L., Linthorst, H. J. M., Bol, J. F. (2001). Role of the 3'-Untranslated Regions of Alfalfa Mosaic Virus RNAs in the Formation of a Transiently Expressed Replicase in Plants and in the Assembly of Virions. J. Virol. 75: 6440-6449 [Abstract] [Full Text]  
  • Huang, C.-Y., Huang, Y.-L., Meng, M., Hsu, Y.-H., Tsai, C.-H. (2001). Sequences at the 3' Untranslated Region of Bamboo Mosaic Potexvirus RNA Interact with the Viral RNA-Dependent RNA Polymerase. J. Virol. 75: 2818-2824 [Abstract] [Full Text]  
  • Li, Y.-I., Chen, Y.-J., Hsu, Y.-H., Meng, M. (2001). Characterization of the AdoMet-Dependent Guanylyltransferase Activity That Is Associated with the N Terminus of Bamboo Mosaic Virus Replicase. J. Virol. 75: 782-788 [Abstract] [Full Text]  
  • Ahola, T., den Boon, J. A., Ahlquist, P. (2000). Helicase and Capping Enzyme Active Site Mutations in Brome Mosaic Virus Protein 1a Cause Defects in Template Recruitment, Negative-Strand RNA Synthesis, and Viral RNA Capping. J. Virol. 74: 8803-8811 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»