Processing of the VP1/2A junction is not necessary for production of foot-and-mouth disease virus empty capsids and infectious viruses: characterization of "self-tagged" particles

Research output: Contribution to journalJournal articlepeer-review

Standard

Processing of the VP1/2A junction is not necessary for production of foot-and-mouth disease virus empty capsids and infectious viruses : characterization of "self-tagged" particles. / Gullberg, Maria; Polacek, Charlotta; Bøtner, Anette; Belsham, Graham J.

In: Journal of Virology, Vol. 87, No. 21, 11.2013, p. 11591-603.

Research output: Contribution to journalJournal articlepeer-review

Harvard

Gullberg, M, Polacek, C, Bøtner, A & Belsham, GJ 2013, 'Processing of the VP1/2A junction is not necessary for production of foot-and-mouth disease virus empty capsids and infectious viruses: characterization of "self-tagged" particles', Journal of Virology, vol. 87, no. 21, pp. 11591-603. https://doi.org/10.1128/JVI.01863-13

APA

Gullberg, M., Polacek, C., Bøtner, A., & Belsham, G. J. (2013). Processing of the VP1/2A junction is not necessary for production of foot-and-mouth disease virus empty capsids and infectious viruses: characterization of "self-tagged" particles. Journal of Virology, 87(21), 11591-603. https://doi.org/10.1128/JVI.01863-13

Vancouver

Gullberg M, Polacek C, Bøtner A, Belsham GJ. Processing of the VP1/2A junction is not necessary for production of foot-and-mouth disease virus empty capsids and infectious viruses: characterization of "self-tagged" particles. Journal of Virology. 2013 Nov;87(21):11591-603. https://doi.org/10.1128/JVI.01863-13

Author

Gullberg, Maria ; Polacek, Charlotta ; Bøtner, Anette ; Belsham, Graham J. / Processing of the VP1/2A junction is not necessary for production of foot-and-mouth disease virus empty capsids and infectious viruses : characterization of "self-tagged" particles. In: Journal of Virology. 2013 ; Vol. 87, No. 21. pp. 11591-603.

Bibtex

@article{d47783e166a240deaf339cf9ee36f2cf,
title = "Processing of the VP1/2A junction is not necessary for production of foot-and-mouth disease virus empty capsids and infectious viruses: characterization of {"}self-tagged{"} particles",
abstract = "The foot-and-mouth disease virus (FMDV) capsid protein precursor, P1-2A, is cleaved by 3C(pro) to generate VP0, VP3, VP1, and the peptide 2A. The capsid proteins self-assemble into empty capsid particles or viruses which do not contain 2A. In a cell culture-adapted strain of FMDV (O1 Manisa [Lindholm]), three different amino acid substitutions (E83K, S134C, and K210E) were identified within the VP1 region of the P1-2A precursor compared to the field strain (wild type [wt]). Expression of the O1 Manisa P1-2A (wt or with the S134C substitution in VP1) plus 3C(pro), using a transient expression system, resulted in efficient capsid protein production and self-assembly of empty capsid particles. Removal of the 2A peptide from the capsid protein precursor had no effect on capsid protein processing or particle assembly. However, modification of E83K alone abrogated particle assembly with no apparent effect on protein processing. Interestingly, the K210E substitution, close to the VP1/2A junction, completely blocked processing by 3C(pro) at this cleavage site, but efficient assembly of {"}self-tagged{"} empty capsid particles, containing the uncleaved VP1-2A, was observed. These self-tagged particles behaved like the unmodified empty capsids in antigen enzyme-linked immunosorbent assays and integrin receptor binding assays. Furthermore, mutant viruses with uncleaved VP1-2A could be rescued in cells from full-length FMDV RNA transcripts encoding the K210E substitution in VP1. Thus, cleavage of the VP1/2A junction is not essential for virus viability. The production of such engineered self-tagged empty capsid particles may facilitate their purification for use as diagnostic reagents and vaccines. ",
keywords = "Amino Acid Substitution, Animals, Capsid/metabolism, Capsid Proteins/genetics, Cell Line, DNA Mutational Analysis, Foot-and-Mouth Disease Virus/genetics, Microbial Viability, Mutant Proteins/genetics, Protein Precursors/genetics, Protein Processing, Post-Translational, Viral Proteins/genetics, Virus Assembly",
author = "Maria Gullberg and Charlotta Polacek and Anette B{\o}tner and Belsham, {Graham J}",
year = "2013",
month = nov,
doi = "10.1128/JVI.01863-13",
language = "English",
volume = "87",
pages = "11591--603",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "21",

}

RIS

TY - JOUR

T1 - Processing of the VP1/2A junction is not necessary for production of foot-and-mouth disease virus empty capsids and infectious viruses

T2 - characterization of "self-tagged" particles

AU - Gullberg, Maria

AU - Polacek, Charlotta

AU - Bøtner, Anette

AU - Belsham, Graham J

PY - 2013/11

Y1 - 2013/11

N2 - The foot-and-mouth disease virus (FMDV) capsid protein precursor, P1-2A, is cleaved by 3C(pro) to generate VP0, VP3, VP1, and the peptide 2A. The capsid proteins self-assemble into empty capsid particles or viruses which do not contain 2A. In a cell culture-adapted strain of FMDV (O1 Manisa [Lindholm]), three different amino acid substitutions (E83K, S134C, and K210E) were identified within the VP1 region of the P1-2A precursor compared to the field strain (wild type [wt]). Expression of the O1 Manisa P1-2A (wt or with the S134C substitution in VP1) plus 3C(pro), using a transient expression system, resulted in efficient capsid protein production and self-assembly of empty capsid particles. Removal of the 2A peptide from the capsid protein precursor had no effect on capsid protein processing or particle assembly. However, modification of E83K alone abrogated particle assembly with no apparent effect on protein processing. Interestingly, the K210E substitution, close to the VP1/2A junction, completely blocked processing by 3C(pro) at this cleavage site, but efficient assembly of "self-tagged" empty capsid particles, containing the uncleaved VP1-2A, was observed. These self-tagged particles behaved like the unmodified empty capsids in antigen enzyme-linked immunosorbent assays and integrin receptor binding assays. Furthermore, mutant viruses with uncleaved VP1-2A could be rescued in cells from full-length FMDV RNA transcripts encoding the K210E substitution in VP1. Thus, cleavage of the VP1/2A junction is not essential for virus viability. The production of such engineered self-tagged empty capsid particles may facilitate their purification for use as diagnostic reagents and vaccines.

AB - The foot-and-mouth disease virus (FMDV) capsid protein precursor, P1-2A, is cleaved by 3C(pro) to generate VP0, VP3, VP1, and the peptide 2A. The capsid proteins self-assemble into empty capsid particles or viruses which do not contain 2A. In a cell culture-adapted strain of FMDV (O1 Manisa [Lindholm]), three different amino acid substitutions (E83K, S134C, and K210E) were identified within the VP1 region of the P1-2A precursor compared to the field strain (wild type [wt]). Expression of the O1 Manisa P1-2A (wt or with the S134C substitution in VP1) plus 3C(pro), using a transient expression system, resulted in efficient capsid protein production and self-assembly of empty capsid particles. Removal of the 2A peptide from the capsid protein precursor had no effect on capsid protein processing or particle assembly. However, modification of E83K alone abrogated particle assembly with no apparent effect on protein processing. Interestingly, the K210E substitution, close to the VP1/2A junction, completely blocked processing by 3C(pro) at this cleavage site, but efficient assembly of "self-tagged" empty capsid particles, containing the uncleaved VP1-2A, was observed. These self-tagged particles behaved like the unmodified empty capsids in antigen enzyme-linked immunosorbent assays and integrin receptor binding assays. Furthermore, mutant viruses with uncleaved VP1-2A could be rescued in cells from full-length FMDV RNA transcripts encoding the K210E substitution in VP1. Thus, cleavage of the VP1/2A junction is not essential for virus viability. The production of such engineered self-tagged empty capsid particles may facilitate their purification for use as diagnostic reagents and vaccines.

KW - Amino Acid Substitution

KW - Animals

KW - Capsid/metabolism

KW - Capsid Proteins/genetics

KW - Cell Line

KW - DNA Mutational Analysis

KW - Foot-and-Mouth Disease Virus/genetics

KW - Microbial Viability

KW - Mutant Proteins/genetics

KW - Protein Precursors/genetics

KW - Protein Processing, Post-Translational

KW - Viral Proteins/genetics

KW - Virus Assembly

U2 - 10.1128/JVI.01863-13

DO - 10.1128/JVI.01863-13

M3 - Journal article

C2 - 23966400

VL - 87

SP - 11591

EP - 11603

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 21

ER -

ID: 257916359