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 Parker, J. S. L.
Right arrow Articles by Nibert, M. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Parker, J. S. L.
Right arrow Articles by Nibert, M. L.

 Previous Article  |  Next Article 

Journal of Virology, May 2002, p. 4483-4496, Vol. 76, No. 9
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.9.4483-4496.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Reovirus Core Protein µ2 Determines the Filamentous Morphology of Viral Inclusion Bodies by Interacting with and Stabilizing Microtubules

John S. L. Parker,1 Teresa J. Broering,1,2 Jonghwa Kim,1,2 Darren E. Higgins,1 and Max L. Nibert1*

Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115,1 Department of Biochemistry, University of Wisconsin—Madison, Madison, Wisconsin 537062

Received 11 October 2001/ Accepted 22 January 2002

Cells infected with mammalian reoviruses often contain large perinuclear inclusion bodies, or "factories," where viral replication and assembly are thought to occur. Here, we report a viral strain difference in the morphology of these inclusions: filamentous inclusions formed in cells infected with reovirus type 1 Lang (T1L), whereas globular inclusions formed in cells infected with our laboratory's isolate of reovirus type 3 Dearing (T3D). Examination by immunofluorescence microscopy revealed the filamentous inclusions to be colinear with microtubules (MTs). The filamentous distribution was dependent on an intact MT network, as depolymerization of MTs early after infection caused globular inclusions to form. The inclusion phenotypes of T1L x T3D reassortant viruses identified the viral M1 genome segment as the primary genetic determinant of the strain difference in inclusion morphology. Filamentous inclusions were seen with 21 of 22 other reovirus strains, including an isolate of T3D obtained from another laboratory. When the µ2 proteins derived from T1L and the other laboratory's T3D isolate were expressed after transfection of their cloned M1 genes, they associated with filamentous structures that colocalized with MTs, whereas the µ2 protein derived from our laboratory's T3D isolate did not. MTs were stabilized in cells infected with the viruses that induced filamentous inclusions and after transfection with the M1 genes derived from those viruses. Evidence for MT stabilization included bundling and hyperacetylation of {alpha}-tubulin, changes characteristically seen when MT-associated proteins (MAPs) are overexpressed. Sequencing of the M1 segments from the different T1L and T3D isolates revealed that a single-amino-acid difference at position 208 correlated with the inclusion morphology. Two mutant forms of µ2 with the changes Pro-208 to Ser in a background of T1L µ2 and Ser-208 to Pro in a background of T3D µ2 had MT association phenotypes opposite to those of the respective wild-type proteins. We conclude that the µ2 protein of most reovirus strains is a viral MAP and that it plays a key role in the formation and structural organization of reovirus inclusion bodies.

* Corresponding author. Mailing address: Department of Microbiology and Molecular Genetics, Harvard Medical School, 200 Longwood Ave., Boston, MA 02115. Phone: (617) 432-4829. Fax: (617) 738-7664. E-mail: mnibert{at}hms.harvard.edu.

Journal of Virology, May 2002, p. 4483-4496, Vol. 76, No. 9
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.9.4483-4496.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Qin, Q., Hastings, C., Miller, C. L. (2009). Mammalian Orthoreovirus Particles Induce and Are Recruited into Stress Granules at Early Times Postinfection. J. Virol. 83: 11090-11101 [Abstract] [Full Text]  
  • Kobayashi, T., Ooms, L. S., Chappell, J. D., Dermody, T. S. (2009). Identification of Functional Domains in Reovirus Replication Proteins {micro}NS and {micro}2. J. Virol. 83: 2892-2906 [Abstract] [Full Text]  
  • Zurney, J., Kobayashi, T., Holm, G. H., Dermody, T. S., Sherry, B. (2009). Reovirus {micro}2 Protein Inhibits Interferon Signaling through a Novel Mechanism Involving Nuclear Accumulation of Interferon Regulatory Factor 9. J. Virol. 83: 2178-2187 [Abstract] [Full Text]  
  • Miller, C. L., Arnold, M. M., Broering, T. J., Eichwald, C., Kim, J., Dinoso, J. B., Nibert, M. L. (2007). Virus-derived Platforms for Visualizing Protein Associations inside Cells. Mol. Cell. Proteomics 6: 1027-1038 [Abstract] [Full Text]  
  • Murray, K. E., Nibert, M. L. (2007). Guanidine Hydrochloride Inhibits Mammalian Orthoreovirus Growth by Reversibly Blocking the Synthesis of Double-Stranded RNA. J. Virol. 81: 4572-4584 [Abstract] [Full Text]  
  • Coffey, C. M., Sheh, A., Kim, I. S., Chandran, K., Nibert, M. L., Parker, J. S. L. (2006). Reovirus Outer Capsid Protein {micro}1 Induces Apoptosis and Associates with Lipid Droplets, Endoplasmic Reticulum, and Mitochondria.. J. Virol. 80: 8422-8438 [Abstract] [Full Text]  
  • Kobayashi, T., Chappell, J. D., Danthi, P., Dermody, T. S. (2006). Gene-specific inhibition of reovirus replication by RNA interference.. J. Virol. 80: 9053-9063 [Abstract] [Full Text]  
  • Warren, J. C., Rutkowski, A., Cassimeris, L. (2006). Infection with Replication-deficient Adenovirus Induces Changes in the Dynamic Instability of Host Cell Microtubules. Mol. Biol. Cell 17: 3557-3568 [Abstract] [Full Text]  
  • Wileman, T. (2006). Aggresomes and autophagy generate sites for virus replication.. Science 312: 875-878 [Abstract] [Full Text]  
  • Wei, T., Shimizu, T., Hagiwara, K., Kikuchi, A., Moriyasu, Y., Suzuki, N., Chen, H., Omura, T. (2006). Pns12 protein of Rice dwarf virus is essential for formation of viroplasms and nucleation of viral-assembly complexes. J. Gen. Virol. 87: 429-438 [Abstract] [Full Text]  
  • Broering, T. J., Arnold, M. M., Miller, C. L., Hurt, J. A., Joyce, P. L., Nibert, M. L. (2005). Carboxyl-Proximal Regions of Reovirus Nonstructural Protein {micro}NS Necessary and Sufficient for Forming Factory-Like Inclusions. J. Virol. 79: 6194-6206 [Abstract] [Full Text]  
  • Ruthel, G., Demmin, G. L., Kallstrom, G., Javid, M. P., Badie, S. S., Will, A. B., Nelle, T., Schokman, R., Nguyen, T. L., Carra, J. H., Bavari, S., Aman, M. J. (2005). Association of Ebola Virus Matrix Protein VP40 with Microtubules. J. Virol. 79: 4709-4719 [Abstract] [Full Text]  
  • Villanueva, R. A., Galaz, J. L., Valdes, J. A., Jashes, M. M., Sandino, A. M. (2004). Genome Assembly and Particle Maturation of the Birnavirus Infectious Pancreatic Necrosis Virus. J. Virol. 78: 13829-13838 [Abstract] [Full Text]  
  • Miller, C. L., Parker, J. S. L., Dinoso, J. B., Piggott, C. D. S., Perron, M. J., Nibert, M. L. (2004). Increased Ubiquitination and Other Covariant Phenotypes Attributed to a Strain- and Temperature-Dependent Defect of Reovirus Core Protein {micro}2. J. Virol. 78: 10291-10302 [Abstract] [Full Text]  
  • Odegard, A. L., Chandran, K., Zhang, X., Parker, J. S. L., Baker, T. S., Nibert, M. L. (2004). Putative Autocleavage of Outer Capsid Protein {micro}1, Allowing Release of Myristoylated Peptide {micro}1N during Particle Uncoating, Is Critical for Cell Entry by Reovirus. J. Virol. 78: 8732-8745 [Abstract] [Full Text]  
  • Jouvenet, N., Monaghan, P., Way, M., Wileman, T. (2004). Transport of African Swine Fever Virus from Assembly Sites to the Plasma Membrane Is Dependent on Microtubules and Conventional Kinesin. J. Virol. 78: 7990-8001 [Abstract] [Full Text]  
  • Hermann, L. L., Coombs, K. M. (2004). Inhibition of Reovirus by Mycophenolic Acid Is Associated with the M1 Genome Segment. J. Virol. 78: 6171-6179 [Abstract] [Full Text]  
  • Nibert, M. L., Kim, J. (2004). Conserved Sequence Motifs for Nucleoside Triphosphate Binding Unique to Turreted Reoviridae Members and Coltiviruses. J. Virol. 78: 5528-5530 [Full Text]  
  • Broering, T. J., Kim, J., Miller, C. L., Piggott, C. D. S., Dinoso, J. B., Nibert, M. L., Parker, J. S. L. (2004). Reovirus Nonstructural Protein {micro}NS Recruits Viral Core Surface Proteins and Entering Core Particles to Factory-Like Inclusions. J. Virol. 78: 1882-1892 [Abstract] [Full Text]  
  • Kim, J., Parker, J. S. L., Murray, K. E., Nibert, M. L. (2004). Nucleoside and RNA Triphosphatase Activities of Orthoreovirus Transcriptase Cofactor {micro}2. J. Biol. Chem. 279: 4394-4403 [Abstract] [Full Text]  
  • Chen, C., Weisz, O. A., Stolz, D. B., Watkins, S. C., Montelaro, R. C. (2004). Differential Effects of Actin Cytoskeleton Dynamics on Equine Infectious Anemia Virus Particle Production. J. Virol. 78: 882-891 [Abstract] [Full Text]  
  • Chandran, K., Parker, J. S. L., Ehrlich, M., Kirchhausen, T., Nibert, M. L. (2003). The {delta} Region of Outer-Capsid Protein {micro}1 Undergoes Conformational Change and Release from Reovirus Particles during Cell Entry. J. Virol. 77: 13361-13375 [Abstract] [Full Text]  
  • Chiou, C.-T., Hu, C.-C. A., Chen, P.-H., Liao, C.-L., Lin, Y.-L., Wang, J.-J. (2003). Association of Japanese encephalitis virus NS3 protein with microtubules and tumour susceptibility gene 101 (TSG101) protein. J. Gen. Virol. 84: 2795-2805 [Abstract] [Full Text]  
  • Becker, M. M., Peters, T. R., Dermody, T. S. (2003). Reovirus {sigma}NS and {micro}NS Proteins Form Cytoplasmic Inclusion Structures in the Absence of Viral Infection. J. Virol. 77: 5948-5963 [Abstract] [Full Text]  
  • Miller, C. L., Broering, T. J., Parker, J. S. L., Arnold, M. M., Nibert, M. L. (2003). Reovirus {sigma}NS Protein Localizes to Inclusions through an Association Requiring the {micro}NS Amino Terminus. J. Virol. 77: 4566-4576 [Abstract] [Full Text]  
  • Heath, C. M., Windsor, M., Wileman, T. (2003). Membrane Association Facilitates the Correct Processing of pp220 during Production of the Major Matrix Proteins of African Swine Fever Virus. J. Virol. 77: 1682-1690 [Abstract] [Full Text]  
  • Broering, T. J., Parker, J. S. L., Joyce, P. L., Kim, J., Nibert, M. L. (2002). Mammalian Reovirus Nonstructural Protein {micro}NS Forms Large Inclusions and Colocalizes with Reovirus Microtubule-Associated Protein {micro}2 in Transfected Cells. J. Virol. 76: 8285-8297 [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»