This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by McAuliffe, J. M. Articles by Skiadopoulos, M. H. Search for Related Content PubMed PubMed Citation Articles by McAuliffe, J. M. Articles by Skiadopoulos, M. H.

 Previous Article  |  Next Article 

Journal of Virology, February 2004, p. 2029-2036, Vol. 78, No. 4
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.4.2029-2036.2004

Codon Substitution Mutations at Two Positions in the L Polymerase Protein of Human Parainfluenza Virus Type 1 Yield Viruses with a Spectrum of Attenuation In Vivo and Increased Phenotypic Stability In Vitro

Josephine M. McAuliffe, Sonja R. Surman, Jason T. Newman, Jeffrey M. Riggs, Peter L. Collins, Brian R. Murphy, and Mario H. Skiadopoulos^*

Respiratory Viruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland

Received 7 July 2003/ Accepted 27 October 2003

The Y942H and L992F temperature-sensitive (ts) and attenuating amino acid substitution mutations, previously identified in the L polymerase of the HPIV3cp45 vaccine candidate, were introduced into homologous positions of the L polymerase of recombinant human parainfluenza virus type 1 (rHPIV1). In rHPIV1, the Y942H mutation specified the ts phenotype in vitro and the attenuation (att) phenotype in hamsters, whereas the L992F mutation specified neither phenotype. Each of these codon mutations was generated by a single nucleotide substitution and therefore had the potential to readily revert to a codon specifying the wild-type amino acid residue. We introduced alternative amino acid assignments at codon 942 or 992 as a strategy to increase genetic stability and to generate mutants that exhibit a range of attenuation. Twenty-three recombinants with codon substitutions at position 942 or 992 of the L protein were viable. One highly ts and att mutant, the Y942A virus, which had a difference of three nucleotides from the codon encoding a wild-type tyrosine, also possessed a high level of genetic and phenotypic stability upon serial passage in vitro at restrictive temperatures compared to that of the parent Y942H virus, which possessed a single nucleotide substitution. We obtained mutants with substitutions at position 992 that, in contrast to the L992F virus, possessed the ts and att phenotypes. These findings identify the use of alternative codon substitution mutations as a method that can be used to generate candidate vaccine viruses with increased genetic stability and/or a modified level of attenuation.

---

* Corresponding author. Mailing address: NIH, Building 50, Room 6511, 50 South Dr., MSC 8007, Bethesda, MD 20892-8007. Phone: (301) 594-2271. Fax: (301) 480-1268. E-mail: mskiadopoulos{at}niaid.nih.gov.

---

Journal of Virology, February 2004, p. 2029-2036, Vol. 78, No. 4
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.4.2029-2036.2004

This article has been cited by other articles:

• Bartlett, E. J., Hennessey, M., Skiadopoulos, M. H., Schmidt, A. C., Collins, P. L., Murphy, B. R., Pickles, R. J. (2008). Role of Interferon in the Replication of Human Parainfluenza Virus Type 1 Wild Type and Mutant Viruses in Human Ciliated Airway Epithelium. J. Virol. 82: 8059-8070 [Abstract] [Full Text]  
• Bukreyev, A., Skiadopoulos, M. H., Murphy, B. R., Collins, P. L. (2006). Nonsegmented negative-strand viruses as vaccine vectors.. J. Virol. 80: 10293-10306 [Full Text]  
• Collins, P. L., Murphy, B. R. (2005). New Generation Live Vaccines against Human Respiratory Syncytial Virus Designed by Reverse Genetics. Proc Am Thorac Soc 2: 166-173 [Abstract] [Full Text]