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 Hwang, L. N.
Right arrow Articles by Pattnaik, A. K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hwang, L. N.
Right arrow Articles by Pattnaik, A. K.

 Previous Article  |  Next Article 

Journal of Virology, July 1999, p. 5613-5620, Vol. 73, No. 7
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Optimal Replication Activity of Vesicular Stomatitis Virus RNA Polymerase Requires Phosphorylation of a Residue(s) at Carboxy-Terminal Domain II of Its Accessory Subunit, Phosphoprotein P

Leroy N. Hwang,1 Nathan Englund,1 Tapas Das,2 Amiya K. Banerjee,2 and Asit K. Pattnaik1,*

Department of Microbiology and Immunology, University of Miami School of Medicine, Miami, Florida 33136,1 and Department of Virology, The Cleveland Clinic Foundation, Cleveland, Ohio 441952

Received 3 September 1998/Accepted 29 March 1999

The phosphoprotein, P, of vesicular stomatitis virus (VSV) is a key subunit of the viral RNA-dependent RNA polymerase complex. The protein is phosphorylated at multiple sites in two different domains. We recently showed that specific serine and threonine residues within the amino-terminal acidic domain I of P protein must be phosphorylated for in vivo transcription activity, but not for replication activity, of the polymerase complex. To examine the role of phosphorylation of the carboxy-terminal domain II residues of the P protein in transcription and replication, we have used a panel of mutant P proteins in which the phosphate acceptor sites (Ser-226, Ser-227, and Ser-233) were altered to alanines either individually or in various combinations. Analyses of the mutant proteins for their ability to support replication of a VSV minigenomic RNA suggest that phosphorylation of either Ser-226 or Ser-227 is necessary for optimal replication activity of the protein. The mutant protein (P226/227) in which both of these residues were altered to alanines was only about 8% active in replication compared to the wild-type (wt) protein. Substitution of alanine for Ser-233 did not have any adverse effect on replication activity of the protein. In contrast, all the mutant proteins showed activities similar to that of the wt protein in transcription. These results indicate that phosphorylation of the carboxy-terminal domain II residues of P protein are required for optimal replication activity but not for transcription activity. Furthermore, substitution of glutamic acid residues for Ser-226 and Ser-227 resulted in a protein that was only 14% active in replication but almost fully active in transcription. Taken together, these results, along with our earlier studies, suggest that phosphorylation of residues at two different domains in the P protein regulates its activity in transcription and replication of the VSV genome.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, University of Miami School of Medicine, P.O. Box 016960 (R-138), Miami, FL 33101. Phone: (305) 243-6711. Fax: (305) 243-4623. E-mail: apattnaik{at}mednet.med.miami.edu.

Journal of Virology, July 1999, p. 5613-5620, Vol. 73, No. 7
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Schmid, S., Mayer, D., Schneider, U., Schwemmle, M. (2007). Functional Characterization of the Major and Minor Phosphorylation Sites of the P Protein of Borna Disease Virus. J. Virol. 81: 5497-5507 [Abstract] [Full Text]  
  • Chen, M., Ogino, T., Banerjee, A. K. (2006). Mapping and Functional Role of the Self-Association Domain of Vesicular Stomatitis Virus Phosphoprotein. J. Virol. 80: 9511-9518 [Abstract] [Full Text]  
  • Das, S. C., Nayak, D., Zhou, Y., Pattnaik, A. K. (2006). Visualization of intracellular transport of vesicular stomatitis virus nucleocapsids in living cells.. J. Virol. 80: 6368-6377 [Abstract] [Full Text]  
  • Ding, H., Green, T. J., Lu, S., Luo, M. (2006). Crystal Structure of the Oligomerization Domain of the Phosphoprotein of Vesicular Stomatitis Virus. J. Virol. 80: 2808-2814 [Abstract] [Full Text]  
  • Dong, B., Zhou, Q., Zhao, J., Zhou, A., Harty, R. N., Bose, S., Banerjee, A., Slee, R., Guenther, J., Williams, B. R. G., Wiedmer, T., Sims, P. J., Silverman, R. H. (2004). Phospholipid Scramblase 1 Potentiates the Antiviral Activity of Interferon. J. Virol. 78: 8983-8993 [Abstract] [Full Text]  
  • Das, S. C., Pattnaik, A. K. (2004). Phosphorylation of Vesicular Stomatitis Virus Phosphoprotein P Is Indispensable for Virus Growth. J. Virol. 78: 6420-6430 [Abstract] [Full Text]  
  • Finke, S., Mueller-Waldeck, R., Conzelmann, K.-K. (2003). Rabies virus matrix protein regulates the balance of virus transcription and replication. J. Gen. Virol. 84: 1613-1621 [Abstract] [Full Text]  
  • Neumann, G., Whitt, M. A., Kawaoka, Y. (2002). A decade after the generation of a negative-sense RNA virus from cloned cDNA - what have we learned?. J. Gen. Virol. 83: 2635-2662 [Abstract] [Full Text]  
  • Lu, B., Ma, C.-H., Brazas, R., Jin, H. (2002). The Major Phosphorylation Sites of the Respiratory Syncytial Virus Phosphoprotein Are Dispensable for Virus Replication In Vitro. J. Virol. 76: 10776-10784 [Abstract] [Full Text]  
  • Goodin, M. M., Austin, J., Tobias, R., Fujita, M., Morales, C., Jackson, A. O. (2001). Interactions and Nuclear Import of the N and P Proteins of Sonchus Yellow Net Virus, a Plant Nucleorhabdovirus. J. Virol. 75: 9393-9406 [Abstract] [Full Text]  
  • Perales, B., Sanz-Ezquerro, J. J., Gastaminza, P., Ortega, J., Santarén, J. F., Ortín, J., Nieto, A. (2000). The Replication Activity of Influenza Virus Polymerase Is Linked to the Capacity of the PA Subunit To Induce Proteolysis. J. Virol. 74: 1307-1312 [Abstract] [Full Text]  
  • Vihinen, H., Ahola, T., Tuittila, M., Merits, A., Kaariainen, L. (2001). Elimination of Phosphorylation Sites of Semliki Forest Virus Replicase Protein nsP3. J. Biol. Chem. 276: 5745-5752 [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»