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PMID: 10074108 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Isolation from tobacco mosaic virus-infected tobacco of a solubilized template-specific RNA-dependent RNA polymerase containing a 126K/183K protein heterodimer.

Journal of virology ·Vol. 73 ·No. 4 ·1999-04-00 ·Pages 2633-40

Watanabe T, Honda A, Iwata A, Ueda S, Hibi T, Ishihama A

Abstract

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.

MeSH Terms
Base Sequence DNA-Directed RNA Polymerases/genetics,metabolism Dimerization Molecular Sequence Data Plants, Toxic RNA, Viral/genetics Recombinant Fusion Proteins/biosynthesis,genetics Templates, Genetic Tobacco/genetics,virology Tobacco Mosaic Virus/genetics Viral Proteins/genetics,metabolism Virus Replication/genetics
Chemicals
RNA, Viral Recombinant Fusion Proteins Viral Proteins DNA-Directed RNA Polymerases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Watanabe T
Department of Molecular Genetics, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan. aishiham@lab.nig.ac.jp
Honda A
Iwata A
Ueda S
Hibi T
Ishihama A
References (31)
31 references, click to expand
  1. In vitro mutagenesis of the putative replicase genes of tobacco mosaic virus.
    Nucleic Acids Res. 1986 Nov 11;14(21):8291-305 PMID: 3786131
  2. Characterization of a host protein associated with brome mosaic virus RNA-dependent RNA polymerase.
    Proc Natl Acad Sci U S A. 1993 Feb 15;90(4):1498-502 PMID: 8434011
  3. Molecular cloning and nucleotide sequence of the 30K and the coat protein cistron of TMV (tomato strain) genome.
    Nucleic Acids Res. 1983 Jun 11;11(11):3767-78 PMID: 6304642
  4. The tobacco mosaic virus RNA polymerase complex contains a plant protein related to the RNA-binding subunit of yeast eIF-3.
    J Virol. 1997 Aug;71(8):6075-82 PMID: 9223501
  5. Complete replication in vitro of tobacco mosaic virus RNA by a template-dependent, membrane-bound RNA polymerase.
    J Virol. 1996 Sep;70(9):6227-34 PMID: 8709249
  6. Purification and characterization of brome mosaic virus RNA-dependent RNA polymerase.
    Virology. 1990 Sep;178(1):189-94 PMID: 2389551
  7. Bacteriophage Q replicase contains the protein biosynthesis elongation factors EF Tu and EF Ts.
    Proc Natl Acad Sci U S A. 1972 May;69(5):1313-7 PMID: 4624757
  8. Promoter analysis of influenza virus RNA polymerase.
    J Virol. 1989 Dec;63(12):5142-52 PMID: 2585601
  9. Identification and sequence determination of the host factor gene for bacteriophage Q beta.
    Nucleic Acids Res. 1991 Mar 11;19(5):1063-6 PMID: 2020545
  10. The rapid isolation of plant virus RNAs using sodium perchlorate.
    J Gen Virol. 1974 Apr;23(1):107-11 PMID: 4364872
  11. Purification of viral RNA by means of bentonite.
    Virology. 1961 May;14:54-8 PMID: 13701154
  12. Functional analysis of deletion mutants of cucumber mosaic virus RNA3 using an in vitro transcription system.
    Virology. 1991 Jul;183(1):106-13 PMID: 2053275
  13. Mutational analysis of the pseudoknot region in the 3' noncoding region of tobacco mosaic virus RNA.
    J Virol. 1990 Aug;64(8):3686-93 PMID: 2370679
  14. The 5'-terminal sequence of TMV RNA. Question on the polymorphism found in vulgare strain.
    FEBS Lett. 1983 Oct 17;162(2):282-5 PMID: 6628671
  15. Cytoplasmic membranes, a possible site of tobacco mosaic virus RNA replication.
    Virology. 1971 Mar;43(3):713-6 PMID: 4107554
  16. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  17. Complete replication of a eukaryotic virus RNA in vitro by a purified RNA-dependent RNA polymerase.
    Cell. 1990 Oct 19;63(2):363-8 PMID: 2208291
  18. Molecular anatomy of viral RNA-directed RNA polymerases.
    Arch Virol. 1994;134(3-4):235-58 PMID: 8129614
  19. Effects of the tom1 mutation of Arabidopsis thaliana on the multiplication of tobacco mosaic virus RNA in protoplasts.
    J Virol. 1993 Sep;67(9):5328-38 PMID: 8350399
  20. Formation of plant RNA virus replication complexes on membranes: role of an endoplasmic reticulum-targeted viral protein.
    EMBO J. 1997 Jul 1;16(13):4049-59 PMID: 9233814
  21. Immunogold localization of the intracellular sites of structural and nonstructural tobacco mosaic virus proteins.
    Virology. 1987 Dec;161(2):488-96 PMID: 3318095
  22. Specific cessation of minus-strand RNA accumulation at an early stage of tobacco mosaic virus infection.
    J Virol. 1991 Feb;65(2):861-8 PMID: 1987377
  23. RNA-dependent replication, transcription, and persistence of brome mosaic virus RNA replicons in S. cerevisiae.
    Cell. 1993 Mar 26;72(6):961-70 PMID: 8458084
  24. Nucleotide sequence and its character of cistron coding for the 30 K protein of tobacco mosaic virus (OM strain).
    J Biochem. 1982 Apr;91(4):1441-4 PMID: 7096297
  25. Terminal uridylyl transferase of Vigna unguiculata: purification and characterization of an enzyme catalyzing the addition of a single UMP residue to the 3'-end of an RNA primer.
    Nucleic Acids Res. 1981 Jun 11;9(11):2433-53 PMID: 6269049
  26. RNA polymerase of influenza virus: role of NP in RNA chain elongation.
    J Biochem. 1988 Dec;104(6):1021-6 PMID: 3243763
  27. Brome mosaic virus helicase- and polymerase-like proteins colocalize on the endoplasmic reticulum at sites of viral RNA synthesis.
    J Virol. 1996 Dec;70(12):8908-16 PMID: 8971020
  28. Subunit I of G beta replicase and 30 S ribosomal protein S1 of Escherichia coli. Evidence for the identity of the two proteins.
    J Biol Chem. 1974 May 25;249(10):3314-6 PMID: 4208476
  29. Formation of brome mosaic virus RNA-dependent RNA polymerase in yeast requires coexpression of viral proteins and viral RNA.
    Proc Natl Acad Sci U S A. 1995 May 23;92(11):4892-6 PMID: 7761419
  30. Isolation of mutants of Arabidopsis thaliana in which accumulation of tobacco mosaic virus coat protein is reduced to low levels.
    Mol Gen Genet. 1991 Nov;230(1-2):33-8 PMID: 1745239
  31. Immunocytochemical localization of the 130K and 180K proteins (putative replicase components) of tobacco mosaic virus.
    Virology. 1987 Oct;160(2):477-81 PMID: 18644575
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.73.4.2633-2640.1999
Email
aishiham@lab.nig.ac.jp
Published
1999-04-00
Pages
2633-40
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC104018
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.73.4.2633-2640.1999 PMC Full Text View journal details
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