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 Restrepo-Hartwig, M.
Right arrow Articles by Ahlquist, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Restrepo-Hartwig, M.
Right arrow Articles by Ahlquist, P.

 Previous Article  |  Next Article 

Journal of Virology, December 1999, p. 10303-10309, Vol. 73, No. 12
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Brome Mosaic Virus RNA Replication Proteins 1a and 2a Colocalize and 1a Independently Localizes on the Yeast Endoplasmic Reticulum

María Restrepo-Hartwig1,dagger and Paul Ahlquist1,2,*

Institute for Molecular Virology1 and Howard Hughes Medical Institute,2 University of Wisconsin---Madison, Madison, Wisconsin 53706

Received 10 June 1999/Accepted 30 August 1999

The universal membrane association of positive-strand RNA virus RNA replication complexes is implicated in their function, but the intracellular membranes used vary among viruses. Brome mosaic virus (BMV) encodes two mutually interacting RNA replication proteins: 1a, which contains RNA capping and helicase-like domains, and the polymerase-like 2a protein. In cells from the natural plant hosts of BMV, 1a and 2a colocalize on the endoplasmic reticulum (ER). 1a and 2a also direct BMV RNA replication and subgenomic mRNA synthesis in the yeast Saccharomyces cerevisiae, but whether the distribution of 1a, 2a, and active replication complexes in yeast duplicates that in plant cells has not been determined. For yeast expressing 1a and 2a and replicating BMV genomic RNA3, we used double-label confocal immunofluorescence to define the localization of 1a, 2a, and viral RNA and to explore the determinants of replication complex targeting. As in plant cells, 1a and 2a colocalized on and were retained on the yeast ER, with no detectable accumulation in the Golgi apparatus. 1a and 2a were distributed over most of the ER surface, with strongest accumulation on the perinuclear ER. In vivo labeling with bromo-UTP showed that the sites of 1a and 2a accumulation were the sites of nascent viral RNA synthesis. In situ hybridization showed that completed viral RNA products accumulated predominantly in the immediate vicinity of replication complexes but that some, possibly more mature cells also accumulated substantial viral RNA in the surrounding cytoplasm distal to replication complexes. Additionally, we find that 1a localizes to the ER when expressed in the absence of other viral factors. These results show that BMV RNA replication in yeast duplicates the normal localization of replication complexes, reveal the intracellular distribution of RNA replication products, and show that 1a is at least partly responsible for the ER localization and retention of the RNA replication complex.

* Corresponding author. Mailing address: Institute for Molecular Virology, University of Wisconsin---Madison, 1525 Linden Dr., Madison, WI 53706-1596. Phone: (608) 263-5916. Fax: (608) 265-9214. E-mail: ahlquist{at}facstaff.wisc.edu.

dagger Present address: DuPont, Wilmington, DE 19880-0015.

Journal of Virology, December 1999, p. 10303-10309, Vol. 73, No. 12
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Sztuba-Solinska, J., Bujarski, J. J. (2008). Insights into the Single-Cell Reproduction Cycle of Members of the Family Bromoviridae: Lessons from the Use of Protoplast Systems. J. Virol. 82: 10330-10340 [Full Text]  
  • Yi, G., Kao, C. (2008). cis- and trans-Acting Functions of Brome Mosaic Virus Protein 1a in Genomic RNA1 Replication. J. Virol. 82: 3045-3053 [Abstract] [Full Text]  
  • Weeks, S. A., Miller, D. J. (2008). The Heat Shock Protein 70 Cochaperone YDJ1 Is Required for Efficient Membrane-Specific Flock House Virus RNA Replication Complex Assembly and Function in Saccharomyces cerevisiae. J. Virol. 82: 2004-2012 [Abstract] [Full Text]  
  • Yi, G., Gopinath, K., Kao, C. C. (2007). Selective Repression of Translation by the Brome Mosaic Virus 1a RNA Replication Protein. J. Virol. 81: 1601-1609 [Abstract] [Full Text]  
  • Ju, H.-J., Brown, J. E., Ye, C.-M., Verchot-Lubicz, J. (2007). Mutations in the Central Domain of Potato Virus X TGBp2 Eliminate Granular Vesicles and Virus Cell-to-Cell Trafficking. J. Virol. 81: 1899-1911 [Abstract] [Full Text]  
  • Navarro, B., Russo, M., Pantaleo, V., Rubino, L. (2006). Cytological analysis of Saccharomyces cerevisiae cells supporting cymbidium ringspot virus defective interfering RNA replication.. J. Gen. Virol. 87: 705-714 [Abstract] [Full Text]  
  • Mas, A., Alves-Rodrigues, I., Noueiry, A., Ahlquist, P., Diez, J. (2006). Host Deadenylation-Dependent mRNA Decapping Factors Are Required for a Key Step in Brome Mosaic Virus RNA Replication. J. Virol. 80: 246-251 [Abstract] [Full Text]  
  • Hwang, M. S., Kim, S. H., Lee, J. H., Bae, J. M., Paek, K. H., Park, Y. I. (2005). Evidence for interaction between the 2a polymerase protein and the 3a movement protein of Cucumber mosaic virus. J. Gen. Virol. 86: 3171-3177 [Abstract] [Full Text]  
  • Wang, X., Lee, W.-M., Watanabe, T., Schwartz, M., Janda, M., Ahlquist, P. (2005). Brome Mosaic Virus 1a Nucleoside Triphosphatase/Helicase Domain Plays Crucial Roles in Recruiting RNA Replication Templates. J. Virol. 79: 13747-13758 [Abstract] [Full Text]  
  • Lukhovitskaya, N. I., Yelina, N. E., Zamyatnin, A. A. Jr, Schepetilnikov, M. V., Solovyev, A. G., Sandgren, M., Morozov, S. Yu., Valkonen, J. P. T., Savenkov, E. I. (2005). Expression, localization and effects on virulence of the cysteine-rich 8 kDa protein of Potato mop-top virus. J. Gen. Virol. 86: 2879-2889 [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]  
  • Grdzelishvili, V. Z., Garcia-Ruiz, H., Watanabe, T., Ahlquist, P. (2005). Mutual Interference between Genomic RNA Replication and Subgenomic mRNA Transcription in Brome Mosaic Virus. J. Virol. 79: 1438-1451 [Abstract] [Full Text]  
  • Moradpour, D., Brass, V., Bieck, E., Friebe, P., Gosert, R., Blum, H. E., Bartenschlager, R., Penin, F., Lohmann, V. (2004). Membrane Association of the RNA-Dependent RNA Polymerase Is Essential for Hepatitis C Virus RNA Replication. J. Virol. 78: 13278-13284 [Abstract] [Full Text]  
  • Schwartz, M., Chen, J., Lee, W.-M., Janda, M., Ahlquist, P. (2004). Alternate, virus-induced membrane rearrangements support positive-strand RNA virus genome replication. Proc. Natl. Acad. Sci. USA 101: 11263-11268 [Abstract] [Full Text]  
  • Pantaleo, V., Rubino, L., Russo, M. (2004). The p36 and p95 replicase proteins of Carnation Italian ringspot virus cooperate in stabilizing defective interfering RNA. J. Gen. Virol. 85: 2429-2433 [Abstract] [Full Text]  
  • Guo, Y. X., Chan, S.-W., Kwang, J. (2004). Membrane Association of Greasy Grouper Nervous Necrosis Virus Protein A and Characterization of Its Mitochondrial Localization Targeting Signal. J. Virol. 78: 6498-6508 [Abstract] [Full Text]  
  • Navarro, B., Rubino, L., Russo, M. (2004). Expression of the Cymbidium Ringspot Virus 33-Kilodalton Protein in Saccharomyces cerevisiae and Molecular Dissection of the Peroxisomal Targeting Signal. J. Virol. 78: 4744-4752 [Abstract] [Full Text]  
  • Hema, M., Kao, C. C. (2004). Template Sequence near the Initiation Nucleotide Can Modulate Brome Mosaic Virus RNA Accumulation in Plant Protoplasts. J. Virol. 78: 1169-1180 [Abstract] [Full Text]  
  • Lee, W.-M., Ahlquist, P. (2003). Membrane Synthesis, Specific Lipid Requirements, and Localized Lipid Composition Changes Associated with a Positive-Strand RNA Virus RNA Replication Protein. J. Virol. 77: 12819-12828 [Abstract] [Full Text]  
  • Miller, D. J., Schwartz, M. D., Dye, B. T., Ahlquist, P. (2003). Engineered Retargeting of Viral RNA Replication Complexes to an Alternative Intracellular Membrane. J. Virol. 77: 12193-12202 [Abstract] [Full Text]  
  • El-Hage, N., Luo, G. (2003). Replication of hepatitis C virus RNA occurs in a membrane-bound replication complex containing nonstructural viral proteins and RNA. J. Gen. Virol. 84: 2761-2769 [Abstract] [Full Text]  
  • Prod'homme, D., Jakubiec, A., Tournier, V., Drugeon, G., Jupin, I. (2003). Targeting of the Turnip Yellow Mosaic Virus 66K Replication Protein to the Chloroplast Envelope Is Mediated by the 140K Protein. J. Virol. 77: 9124-9135 [Abstract] [Full Text]  
  • Noueiry, A. O., Diez, J., Falk, S. P., Chen, J., Ahlquist, P. (2003). Yeast Lsm1p-7p/Pat1p Deadenylation-Dependent mRNA-Decapping Factors Are Required for Brome Mosaic Virus Genomic RNA Translation. Mol. Cell. Biol. 23: 4094-4106 [Abstract] [Full Text]  
  • Tomita, Y., Mizuno, T., Diez, J., Naito, S., Ahlquist, P., Ishikawa, M. (2003). Mutation of Host dnaJ Homolog Inhibits Brome Mosaic Virus Negative-Strand RNA Synthesis. J. Virol. 77: 2990-2997 [Abstract] [Full Text]  
  • Chen, J., Noueiry, A., Ahlquist, P. (2003). An Alternate Pathway for Recruiting Template RNA to the Brome Mosaic Virus RNA Replication Complex. J. Virol. 77: 2568-2577 [Abstract] [Full Text]  
  • Pantaleo, V., Rubino, L., Russo, M. (2003). Replication of Carnation Italian Ringspot Virus Defective Interfering RNA in Saccharomyces cerevisiae. J. Virol. 77: 2116-2123 [Abstract] [Full Text]  
  • Dohi, K., Mise, K., Furusawa, I., Okuno, T. (2002). RNA-dependent RNA polymerase complex of Brome mosaic virus: analysis of the molecular structure with monoclonal antibodies. J. Gen. Virol. 83: 2879-2890 [Abstract] [Full Text]  
  • Vlot, A. C., Menard, A., Bol, J. F. (2002). Role of the Alfalfa Mosaic Virus Methyltransferase-Like Domain in Negative-Strand RNA Synthesis. J. Virol. 76: 11321-11328 [Abstract] [Full Text]  
  • Miller, D. J., Ahlquist, P. (2002). Flock House Virus RNA Polymerase Is a Transmembrane Protein with Amino-Terminal Sequences Sufficient for Mitochondrial Localization and Membrane Insertion. J. Virol. 76: 9856-9867 [Abstract] [Full Text]  
  • Ritzenthaler, C., Laporte, C., Gaire, F., Dunoyer, P., Schmitt, C., Duval, S., Piequet, A., Loudes, A. M., Rohfritsch, O., Stussi-Garaud, C., Pfeiffer, P. (2002). Grapevine Fanleaf Virus Replication Occurs on Endoplasmic Reticulum-Derived Membranes. J. Virol. 76: 8808-8819 [Abstract] [Full Text]  
  • Dunoyer, P., Ritzenthaler, C., Hemmer, O., Michler, P., Fritsch, C. (2002). Intracellular Localization of the Peanut Clump Virus Replication Complex in Tobacco BY-2 Protoplasts Containing Green Fluorescent Protein-Labeled Endoplasmic Reticulum or Golgi Apparatus. J. Virol. 76: 865-874 [Abstract] [Full Text]  
  • den Boon, J. A., Chen, J., Ahlquist, P. (2001). Identification of Sequences in Brome Mosaic Virus Replicase Protein 1a That Mediate Association with Endoplasmic Reticulum Membranes. J. Virol. 75: 12370-12381 [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]  
  • Chen, J., Noueiry, A., Ahlquist, P. (2001). Brome Mosaic Virus Protein 1a Recruits Viral RNA2 to RNA Replication through a 5' Proximal RNA2 Signal. J. Virol. 75: 3207-3219 [Abstract] [Full Text]  
  • Lee, W.-M., Ishikawa, M., Ahlquist, P. (2001). Mutation of Host {Delta}9 Fatty Acid Desaturase Inhibits Brome Mosaic Virus RNA Replication between Template Recognition and RNA Synthesis. J. Virol. 75: 2097-2106 [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]  
  • Dunoyer, P., Herzog, E., Hemmer, O., Ritzenthaler, C., Fritsch, C. (2001). Peanut Clump Virus RNA-1-Encoded P15 Regulates Viral RNA Accumulation but Is Not Abundant at Viral RNA Replication Sites. J. Virol. 75: 1941-1948 [Abstract] [Full Text]  
  • Noueiry, A. O., Chen, J., Ahlquist, P. (2000). A mutant allele of essential, general translation initiation factor DED1 selectively inhibits translation of a viral mRNA. Proc. Natl. Acad. Sci. USA 10.1073/pnas.240460897v1 [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]  
  • Ishikawa, M., Janda, M., Ahlquist, P. (2000). The 3a cell-to-cell movement gene is dispensable for cell-to-cell transmission of brome mosaic virus RNA replicons in yeast but retained over 1045-fold amplification. J. Gen. Virol. 81: 2307-2311 [Abstract] [Full Text]  
  • Carette, J. E., Stuiver, M., Van Lent, J., Wellink, J., Van Kammen, A. (2000). Cowpea Mosaic Virus Infection Induces a Massive Proliferation of Endoplasmic Reticulum but Not Golgi Membranes and Is Dependent on De Novo Membrane Synthesis. J. Virol. 74: 6556-6563 [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]  
  • Ahola, T., Ahlquist, P. (1999). Putative RNA Capping Activities Encoded by Brome Mosaic Virus: Methylation and Covalent Binding of Guanylate by Replicase Protein 1a. J. Virol. 73: 10061-10069 [Abstract] [Full Text]  
  • Diez, J., Ishikawa, M., Kaido, M., Ahlquist, P. (2000). Identification and characterization of a host protein required for efficient template selection in viral RNA replication. Proc. Natl. Acad. Sci. USA 97: 3913-3918 [Abstract] [Full Text]  
  • Noueiry, A. O., Chen, J., Ahlquist, P. (2000). A mutant allele of essential, general translation initiation factor DED1 selectively inhibits translation of a viral mRNA. Proc. Natl. Acad. Sci. USA 97: 12985-12990 [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»