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 Watanabe, T.
Right arrow Articles by Ishihama, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Watanabe, T.
Right arrow Articles by Ishihama, A.

 Previous Article  |  Next Article 

Journal of Virology, April 1999, p. 2633-2640, Vol. 73, No. 4
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Isolation from Tobacco Mosaic Virus-Infected Tobacco of a Solubilized Template-Specific RNA-Dependent RNA Polymerase Containing a 126K/183K Protein Heterodimer

Takato Watanabe,1,2 Ayae Honda,1 Akira Iwata,3 Susumu Ueda,3 Tadaaki Hibi,2 and Akira Ishihama1,*

Department of Molecular Genetics, National Institute of Genetics, Mishima, Shizuoka 411-8540,1 Department of Agricultural and Environmental Biology, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657,2 and Nippon Institute for Biological Science, Ohme, Tokyo 198-0024,3 Japan

Received 19 October 1998/Accepted 16 December 1998

The complete nucleotide sequence was determined for the putative RNA polymerase (183K protein) gene of tobacco mosaic virus (TMV) OM strain, which differed from the related strain, vulgare, by 51 positions in its nucleotide sequence and 6 residues in its amino acid sequence. Three segments of this 183K protein, each containing the sequence motif of methyltransferase (M), helicase (H), or RNA-dependent RNA polymerase (P), were expressed in Escherichia coli as fusion proteins with hexahistidine tags, and domain-specific antibodies were raised against purified His-tagged M and P polypeptides. By immunoaffinity purification, a template-specific RNA-dependent RNA polymerase containing a heterodimer of the full-length 183K and 126K (an amino-terminal-proximal portion of the 183K protein) viral proteins was isolated. We propose that the TMV RNA polymerase for minus-strand RNA synthesis is composed of one molecule each of the 183- and 126-kDa proteins, possibly together with two or more host proteins.


* Corresponding author. Mailing address: Department of Molecular Genetics, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan. Phone: 81-559-81-6741. Fax: 81-559-81-6746. E-mail: aishiham{at}lab.nig.ac.jp.


Journal of Virology, April 1999, p. 2633-2640, Vol. 73, No. 4
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.



This article has been cited by other articles:

  • Csorba, T., Bovi, A., Dalmay, T., Burgyan, J. (2007). The p122 Subunit of Tobacco Mosaic Virus Replicase Is a Potent Silencing Suppressor and Compromises both Small Interfering RNA- and MicroRNA-Mediated Pathways. J. Virol. 81: 11768-11780 [Abstract] [Full Text]  
  • Vogler, H., Akbergenov, R., Shivaprasad, P. V., Dang, V., Fasler, M., Kwon, M.-O., Zhanybekova, S., Hohn, T., Heinlein, M. (2007). Modification of Small RNAs Associated with Suppression of RNA Silencing by Tobamovirus Replicase Protein. J. Virol. 81: 10379-10388 [Abstract] [Full Text]  
  • Komoda, K., Mawatari, N., Hagiwara-Komoda, Y., Naito, S., Ishikawa, M. (2007). Identification of a Ribonucleoprotein Intermediate of Tomato Mosaic Virus RNA Replication Complex Formation. J. Virol. 81: 2584-2591 [Abstract] [Full Text]  
  • Nishikiori, M., Dohi, K., Mori, M., Meshi, T., Naito, S., Ishikawa, M. (2006). Membrane-bound tomato mosaic virus replication proteins participate in RNA synthesis and are associated with host proteins in a pattern distinct from those that are not membrane bound.. J. Virol. 80: 8459-8468 [Abstract] [Full Text]  
  • Liu, J.-Z., Blancaflor, E. B., Nelson, R. S. (2005). The Tobacco Mosaic Virus 126-Kilodalton Protein, a Constituent of the Virus Replication Complex, Alone or within the Complex Aligns with and Traffics along Microfilaments. Plant Physiol. 138: 1853-1865 [Abstract] [Full Text]  
  • Canto, T., MacFarlane, S. A., Palukaitis, P. (2004). ORF6 of Tobacco mosaic virus is a determinant of viral pathogenicity in Nicotiana benthamiana. J. Gen. Virol. 85: 3123-3133 [Abstract] [Full Text]  
  • Panaviene, Z., Panavas, T., Serva, S., Nagy, P. D. (2004). Purification of the Cucumber Necrosis Virus Replicase from Yeast Cells: Role of Coexpressed Viral RNA in Stimulation of Replicase Activity. J. Virol. 78: 8254-8263 [Abstract] [Full Text]  
  • Wang, H.-H., Wong, S.-M. (2004). Significance of the 3'-terminal region in minus-strand RNA synthesis of Hibiscus chlorotic ringspot virus. J. Gen. Virol. 85: 1763-1776 [Abstract] [Full Text]  
  • Hirashima, K., Watanabe, Y. (2003). RNA Helicase Domain of Tobamovirus Replicase Executes Cell-to-Cell Movement Possibly through Collaboration with Its Nonconserved Region. J. Virol. 77: 12357-12362 [Abstract] [Full Text]  
  • Vlot, A. C., Laros, S. M., Bol, J. F. (2003). Coordinate Replication of Alfalfa Mosaic Virus RNAs 1 and 2 Involves cis- and trans-Acting Functions of the Encoded Helicase-Like and Polymerase-Like Domains. J. Virol. 77: 10790-10798 [Abstract] [Full Text]  
  • Osman, T. A. M., Buck, K. W. (2003). Identification of a Region of the Tobacco Mosaic Virus 126- and 183-Kilodalton Replication Proteins Which Binds Specifically to the Viral 3'-Terminal tRNA-Like Structure. J. Virol. 77: 8669-8675 [Abstract] [Full Text]  
  • Goregaoker, S. P., Culver, J. N. (2003). Oligomerization and Activity of the Helicase Domain of the Tobacco Mosaic Virus 126- and 183-Kilodalton Replicase Proteins. J. Virol. 77: 3549-3556 [Abstract] [Full Text]  
  • Shirako, Y., Strauss, E. G., Strauss, J. H. (2003). Modification of the 5' Terminus of Sindbis Virus Genomic RNA Allows nsP4 RNA Polymerases with Nonaromatic Amino Acids at the N Terminus To Function in RNA Replication. J. Virol. 77: 2301-2309 [Abstract] [Full Text]  
  • Zeenko, V. V., Ryabova, L. A., Spirin, A. S., Rothnie, H. M., Hess, D., Browning, K. S., Hohn, T. (2002). Eukaryotic Elongation Factor 1A Interacts with the Upstream Pseudoknot Domain in the 3' Untranslated Region of Tobacco Mosaic Virus RNA. J. Virol. 76: 5678-5691 [Abstract] [Full Text]  
  • Yamanaka, T., Imai, T., Satoh, R., Kawashima, A., Takahashi, M., Tomita, K., Kubota, K., Meshi, T., Naito, S., Ishikawa, M. (2002). Complete Inhibition of Tobamovirus Multiplication by Simultaneous Mutations in Two Homologous Host Genes. J. Virol. 76: 2491-2497 [Abstract] [Full Text]  
  • Knapp, E., Dawson, W. O., Lewandowski, D. J. (2001). Conundrum of the Lack of Defective RNAs (dRNAs) Associated with Tobamovirus Infections: dRNAs That Can Move Are Not Replicated by the Wild-Type Virus; dRNAs That Are Replicated by the Wild-Type Virus Do Not Move. J. Virol. 75: 5518-5525 [Abstract] [Full Text]  
  • Van Der Heijden, M. W., Carette, J. E., Reinhoud, P. J., Haegi, A., Bol, J. F. (2001). Alfalfa Mosaic Virus Replicase Proteins P1 and P2 Interact and Colocalize at the Vacuolar Membrane. J. Virol. 75: 1879-1887 [Abstract] [Full Text]  
  • Osman, T. A. M., Hemenway, C. L., Buck, K. W. (2000). Role of the 3' tRNA-Like Structure in Tobacco Mosaic Virus Minus-Strand RNA Synthesis by the Viral RNA-Dependent RNA Polymerase In Vitro. J. Virol. 74: 11671-11680 [Abstract] [Full Text]  
  • Taylor, D. N., Carr, J. P. (2000). The GCD10 subunit of yeast eIF-3 binds the methyltransferase-like domain of the 126 and 183 kDa replicase proteins of tobacco mosaic virus in the yeast two-hybrid system. J. Gen. Virol. 81: 1587-1591 [Abstract] [Full Text]  
  • Chen, J., Ahlquist, P. (2000). Brome Mosaic Virus Polymerase-Like Protein 2a Is Directed to the Endoplasmic Reticulum by Helicase-Like Viral Protein 1a. J. Virol. 74: 4310-4318 [Abstract] [Full Text]