Determination of Spontaneous Mutation Frequencies in Measles Virus under Nonselective Conditions

Xiaomeng Zhang, Linda J. Rennick, W. Paul Duprex, Bert K. Rima*

*Corresponding author for this work

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

There is a paradox between the remarkable genetic stability of measles virus (MV) in the field and the high mutation rates implied by the frequency of the appearance of monoclonal antibody escape mutants generated when the virus is pressured to revert in vitro (S. J. Schrag, P. A. Rota, and W. J. Bellini, J. Virol. 73: 51-54, 1999). We established a highly sensitive assay to determine frequencies of various categories of mutations in large populations of wild-type and laboratory-adapted MVs using recombinant viruses containing an additional transcription unit (ATU) encoding enhanced green fluorescent protein (EGFP). Single and double mutations were made in the fluorophore of EGFP to ablate fluorescence. The frequencies of reversion mutants in the population were determined by measuring the appearance of fluorescence indicating a revertant virus. This allows mutation rates to be measured under nonselective conditions, as phenotypic reversion to fluorescence requires only either a single-or a double-nucleotide change and amino acid substitution, which does not affect the length of the nonessential reporter protein expressed from the ATU. Mutation rates in MV are the same for wild-type and laboratory-adapted viruses, and they are an order of magnitude lower than the previous measurement assessed under selective conditions. The actual mutation rate for MV is approximately 1.8 x 10(-6) per base per replication event. Copyright © 2013, American Society for Microbiology. All Rights Reserved.

Original languageEnglish
Pages (from-to)2686-2692
Number of pages7
JournalJournal of Virology
Volume87
Issue number5
DOIs
Publication statusPublished - Mar 2013

Keywords

  • RNA
  • HEMAGGLUTININ
  • INFECTIONS
  • DIVERGENCE
  • SELECTION
  • SEQUENCE

Cite this

Zhang, Xiaomeng ; Rennick, Linda J. ; Duprex, W. Paul ; Rima, Bert K. / Determination of Spontaneous Mutation Frequencies in Measles Virus under Nonselective Conditions. In: Journal of Virology. 2013 ; Vol. 87, No. 5. pp. 2686-2692.
@article{8551546b4054424d88846e55645a4a47,
title = "Determination of Spontaneous Mutation Frequencies in Measles Virus under Nonselective Conditions",
abstract = "There is a paradox between the remarkable genetic stability of measles virus (MV) in the field and the high mutation rates implied by the frequency of the appearance of monoclonal antibody escape mutants generated when the virus is pressured to revert in vitro (S. J. Schrag, P. A. Rota, and W. J. Bellini, J. Virol. 73: 51-54, 1999). We established a highly sensitive assay to determine frequencies of various categories of mutations in large populations of wild-type and laboratory-adapted MVs using recombinant viruses containing an additional transcription unit (ATU) encoding enhanced green fluorescent protein (EGFP). Single and double mutations were made in the fluorophore of EGFP to ablate fluorescence. The frequencies of reversion mutants in the population were determined by measuring the appearance of fluorescence indicating a revertant virus. This allows mutation rates to be measured under nonselective conditions, as phenotypic reversion to fluorescence requires only either a single-or a double-nucleotide change and amino acid substitution, which does not affect the length of the nonessential reporter protein expressed from the ATU. Mutation rates in MV are the same for wild-type and laboratory-adapted viruses, and they are an order of magnitude lower than the previous measurement assessed under selective conditions. The actual mutation rate for MV is approximately 1.8 x 10(-6) per base per replication event. Copyright {\circledC} 2013, American Society for Microbiology. All Rights Reserved.",
keywords = "RNA, HEMAGGLUTININ, INFECTIONS, DIVERGENCE, SELECTION, SEQUENCE",
author = "Xiaomeng Zhang and Rennick, {Linda J.} and Duprex, {W. Paul} and Rima, {Bert K.}",
year = "2013",
month = "3",
doi = "10.1128/JVI.02146-12",
language = "English",
volume = "87",
pages = "2686--2692",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "5",

}

Determination of Spontaneous Mutation Frequencies in Measles Virus under Nonselective Conditions. / Zhang, Xiaomeng; Rennick, Linda J.; Duprex, W. Paul; Rima, Bert K.

In: Journal of Virology, Vol. 87, No. 5, 03.2013, p. 2686-2692.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Determination of Spontaneous Mutation Frequencies in Measles Virus under Nonselective Conditions

AU - Zhang, Xiaomeng

AU - Rennick, Linda J.

AU - Duprex, W. Paul

AU - Rima, Bert K.

PY - 2013/3

Y1 - 2013/3

N2 - There is a paradox between the remarkable genetic stability of measles virus (MV) in the field and the high mutation rates implied by the frequency of the appearance of monoclonal antibody escape mutants generated when the virus is pressured to revert in vitro (S. J. Schrag, P. A. Rota, and W. J. Bellini, J. Virol. 73: 51-54, 1999). We established a highly sensitive assay to determine frequencies of various categories of mutations in large populations of wild-type and laboratory-adapted MVs using recombinant viruses containing an additional transcription unit (ATU) encoding enhanced green fluorescent protein (EGFP). Single and double mutations were made in the fluorophore of EGFP to ablate fluorescence. The frequencies of reversion mutants in the population were determined by measuring the appearance of fluorescence indicating a revertant virus. This allows mutation rates to be measured under nonselective conditions, as phenotypic reversion to fluorescence requires only either a single-or a double-nucleotide change and amino acid substitution, which does not affect the length of the nonessential reporter protein expressed from the ATU. Mutation rates in MV are the same for wild-type and laboratory-adapted viruses, and they are an order of magnitude lower than the previous measurement assessed under selective conditions. The actual mutation rate for MV is approximately 1.8 x 10(-6) per base per replication event. Copyright © 2013, American Society for Microbiology. All Rights Reserved.

AB - There is a paradox between the remarkable genetic stability of measles virus (MV) in the field and the high mutation rates implied by the frequency of the appearance of monoclonal antibody escape mutants generated when the virus is pressured to revert in vitro (S. J. Schrag, P. A. Rota, and W. J. Bellini, J. Virol. 73: 51-54, 1999). We established a highly sensitive assay to determine frequencies of various categories of mutations in large populations of wild-type and laboratory-adapted MVs using recombinant viruses containing an additional transcription unit (ATU) encoding enhanced green fluorescent protein (EGFP). Single and double mutations were made in the fluorophore of EGFP to ablate fluorescence. The frequencies of reversion mutants in the population were determined by measuring the appearance of fluorescence indicating a revertant virus. This allows mutation rates to be measured under nonselective conditions, as phenotypic reversion to fluorescence requires only either a single-or a double-nucleotide change and amino acid substitution, which does not affect the length of the nonessential reporter protein expressed from the ATU. Mutation rates in MV are the same for wild-type and laboratory-adapted viruses, and they are an order of magnitude lower than the previous measurement assessed under selective conditions. The actual mutation rate for MV is approximately 1.8 x 10(-6) per base per replication event. Copyright © 2013, American Society for Microbiology. All Rights Reserved.

KW - RNA

KW - HEMAGGLUTININ

KW - INFECTIONS

KW - DIVERGENCE

KW - SELECTION

KW - SEQUENCE

U2 - 10.1128/JVI.02146-12

DO - 10.1128/JVI.02146-12

M3 - Article

VL - 87

SP - 2686

EP - 2692

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 5

ER -