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 Mallardo, M.
Right arrow Articles by Krijnse Locker, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mallardo, M.
Right arrow Articles by Krijnse Locker, J.

 Previous Article  |  Next Article 

Journal of Virology, May 2002, p. 5167-5183, Vol. 76, No. 10
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.10.5167-5183.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Relationship between Vaccinia Virus Intracellular Cores, Early mRNAs, and DNA Replication Sites

Massimo Mallardo,1,{dagger} Edward Leithe,2 Sibylle Schleich,1 Norbert Roos,2 Laura Doglio,1 and Jacomine Krijnse Locker1*

Cell Biology and Biophysics Programme, European Molecular Biology Laboratory, 69117 Heidelberg, Germany,1 Electron Microscopy Unit for Biological Sciences, University of Oslo, Blindren, Oslo, Norway2

Received 4 September 2001/ Accepted 15 February 2002

Virus assembly, a late event in the life cycle of vaccinia virus (VV), is preceded by a number of steps that all occur in the cytoplasm of the infected host cell: virion entry, delivery of the viral core into the cytoplasm, and transcription from these cores of early mRNAs, followed by the process of DNA replication. In the present study the quantitative and structural relationships between these distinct steps of VV morphogenesis were investigated. We show that viral RNA and DNA synthesis increases linearly with increasing amounts of incoming cores. Moreover, at multiplicities of infection that result in 10 to 40 cores per cell, an approximately 1:1 ratio between cores and sites of DNA replication exists, suggesting that each core is infectious. We have shown previously that VV early mRNAs collect in distinct granular structures that recruit components of the host cell translation machinery. Strikingly, these structures appeared to form some distance away from intracellular cores (M. Mallardo, S. Schleich, and J. Krijnse Locker, Mol. Biol. Cell 12:3875-3891, 2001). In the present study the intracellular locations of the sites of early mRNA accumulation and those of the subsequent process of DNA replication were compared. We show that these are distinct structures that have different intracellular locations. Finally, we study the fate of the parental DNA after core uncoating. By electron microscopy, cores were found close to membranes of the endoplasmic reticulum (ER) and the parental DNA, once it had left the core, appeared to associate preferentially with the cytosolic side of those membranes. Since we have previously shown that the process of DNA replication occurs in an ER-enclosed cytosolic "subcompartment" (N. Tolonen, L. Doglio, S. Schleich, and J. Krijnse Locker, Mol. Biol. Cell 12:2031-2046, 2001), the present data suggest that the parental DNA is released into the cytosol and associates with the same membranes where DNA replication is subsequently initiated. The combined data are discussed with respect to the cytosolic organization of VV morphogenesis.

* Corresponding author. Mailing address: EMBL, Meyerhofstrasse 1, 69117 Heidelberg, Germany. Phone: 49 6221 387 508. Fax: 49 6221 387 306. E-mail: Krijnse{at}EMBL-Heidelberg.de.

{dagger} Present address: Max Planck Institute for Developmental Biology, 72076 Tuebingen, Germany.

Journal of Virology, May 2002, p. 5167-5183, Vol. 76, No. 10
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.10.5167-5183.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Cyrklaff, M., Risco, C., Fernandez, J. J., Jimenez, M. V., Esteban, M., Baumeister, W., Carrascosa, J. L. (2005). Cryo-electron tomography of vaccinia virus. Proc. Natl. Acad. Sci. USA 102: 2772-2777 [Abstract] [Full Text]  
  • Welsch, S., Doglio, L., Schleich, S., Krijnse Locker, J. (2003). The Vaccinia Virus I3L Gene Product Is Localized to a Complex Endoplasmic Reticulum-Associated Structure That Contains the Viral Parental DNA. J. Virol. 77: 6014-6028 [Abstract] [Full Text]  
  • Doglio, L., De Marco, A., Schleich, S., Roos, N., Krijnse Locker, J. (2002). The Vaccinia Virus E8R Gene Product: a Viral Membrane Protein That Is Made Early in Infection and Packaged into the Virions' Core. J. Virol. 76: 9773-9786 [Abstract] [Full Text]  
  • Sancho, M. C., Schleich, S., Griffiths, G., Krijnse-Locker, J. (2002). The Block in Assembly of Modified Vaccinia Virus Ankara in HeLa Cells Reveals New Insights into Vaccinia Virus Morphogenesis. J. Virol. 76: 8318-8334 [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»