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Interactome analysis of the human respiratory syncytial virus RNA polymerase complex identifies protein chaperones as important co-factors that promote L protein stability and RNA synthesis

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Interactome analysis of the human respiratory syncytial virus RNA polymerase complex identifies protein chaperones as important co-factors that promote L protein stability and RNA synthesis. / Munday, Diane Carolyn; Wu, Weining; Smith, Nikki; Fix, Jenna; Noton, Sarah Louise; Galloux, Marie; Touzelet, Olivier; Armstrong, Stuart D; Dawson, Jenna M; Aljabr, Waleed; Easton, Andrew J; Rameix-Welti, Marie-Anne; de Oliveira, Andressa Peres; Simabuco, Fernando; Ventura, Armando M; Hughes, David J; Barr, John N; Fearns, Rachel; Digard, Paul; Eléouët, Jean-François; Hiscox, Julian A.

In: Journal of Virology, Vol. 89, No. 2, 15.01.2015, p. 917-930.

Research output: Contribution to journalArticlepeer-review

Harvard

Munday, DC, Wu, W, Smith, N, Fix, J, Noton, SL, Galloux, M, Touzelet, O, Armstrong, SD, Dawson, JM, Aljabr, W, Easton, AJ, Rameix-Welti, M-A, de Oliveira, AP, Simabuco, F, Ventura, AM, Hughes, DJ, Barr, JN, Fearns, R, Digard, P, Eléouët, J-F & Hiscox, JA 2015, 'Interactome analysis of the human respiratory syncytial virus RNA polymerase complex identifies protein chaperones as important co-factors that promote L protein stability and RNA synthesis', Journal of Virology, vol. 89, no. 2, pp. 917-930. https://doi.org/10.1128/JVI.01783-14

APA

Munday, D. C., Wu, W., Smith, N., Fix, J., Noton, S. L., Galloux, M., Touzelet, O., Armstrong, S. D., Dawson, J. M., Aljabr, W., Easton, A. J., Rameix-Welti, M-A., de Oliveira, A. P., Simabuco, F., Ventura, A. M., Hughes, D. J., Barr, J. N., Fearns, R., Digard, P., ... Hiscox, J. A. (2015). Interactome analysis of the human respiratory syncytial virus RNA polymerase complex identifies protein chaperones as important co-factors that promote L protein stability and RNA synthesis. Journal of Virology, 89(2), 917-930. https://doi.org/10.1128/JVI.01783-14

Vancouver

Munday DC, Wu W, Smith N, Fix J, Noton SL, Galloux M et al. Interactome analysis of the human respiratory syncytial virus RNA polymerase complex identifies protein chaperones as important co-factors that promote L protein stability and RNA synthesis. Journal of Virology. 2015 Jan 15;89(2):917-930. https://doi.org/10.1128/JVI.01783-14

Author

Munday, Diane Carolyn ; Wu, Weining ; Smith, Nikki ; Fix, Jenna ; Noton, Sarah Louise ; Galloux, Marie ; Touzelet, Olivier ; Armstrong, Stuart D ; Dawson, Jenna M ; Aljabr, Waleed ; Easton, Andrew J ; Rameix-Welti, Marie-Anne ; de Oliveira, Andressa Peres ; Simabuco, Fernando ; Ventura, Armando M ; Hughes, David J ; Barr, John N ; Fearns, Rachel ; Digard, Paul ; Eléouët, Jean-François ; Hiscox, Julian A. / Interactome analysis of the human respiratory syncytial virus RNA polymerase complex identifies protein chaperones as important co-factors that promote L protein stability and RNA synthesis. In: Journal of Virology. 2015 ; Vol. 89, No. 2. pp. 917-930.

Bibtex - Download

@article{47f3ee5c1a5a42498a8141214565190b,
title = "Interactome analysis of the human respiratory syncytial virus RNA polymerase complex identifies protein chaperones as important co-factors that promote L protein stability and RNA synthesis",
abstract = "The human respiratory syncytial virus (HRSV) core viral RNA polymerase comprises the large polymerase protein (L) and its co-factor the phosphoprotein (P) which associate with the viral ribonucleoprotein complex to replicate the genome and, together with the M2-1 protein, transcribe viral mRNAs. Whilst cellular proteins have long been proposed to be involved in the synthesis of HRSV RNA by associating with the polymerase complex, their characterization has been hindered by the difficulty of purifying the viral polymerase from mammalian cell culture. In this study, EGFP-tagged L and P protein expression was coupled with high affinity anti-GFP antibody-based immunoprecipitation and quantitative proteomics to identify cellular proteins that interacted with either the L or the P proteins when expressed as part of a biologically active viral RNP. Several core groups of cellular proteins were identified that interacted with each viral protein, including in both cases, protein chaperones. Ablation of chaperone activity using small molecule inhibitors confirmed previous studies, which suggested this class of proteins acted as positive viral factors. Inhibition of HSP90 chaperone function in the current study showed that HSP90 was critical for L protein function and stability, whether in the presence or absence of the P protein. Inhibition studies suggested that HSP70 also disrupted virus biology and might help the polymerase remodel the nucleocapsid to allow RNA synthesis to occur efficiently. This indicated a pro-viral role for protein chaperones in HRSV replication and demonstrates that the function of cellular proteins can be targeted as potential therapeutics to disrupt virus replication.",
author = "Munday, {Diane Carolyn} and Weining Wu and Nikki Smith and Jenna Fix and Noton, {Sarah Louise} and Marie Galloux and Olivier Touzelet and Armstrong, {Stuart D} and Dawson, {Jenna M} and Waleed Aljabr and Easton, {Andrew J} and Marie-Anne Rameix-Welti and {de Oliveira}, {Andressa Peres} and Fernando Simabuco and Ventura, {Armando M} and Hughes, {David J} and Barr, {John N} and Rachel Fearns and Paul Digard and Jean-Fran{\c c}ois El{\'e}ou{\"e}t and Hiscox, {Julian A}",
note = "This research was supported by the award of a Medical Research Council (MRC) Project Grant (MR/K000276/1) to P.D., J.N.B., and J.A.H., an MRC studentship to J.N.B. and J.A.H., NIHR funding to J.A.H., and NIH grant R01AI074903 to R.F. W.A. is supported by the Ministry of Education, Kingdom of Saudi Arabia. ",
year = "2015",
month = jan,
day = "15",
doi = "10.1128/JVI.01783-14",
language = "English",
volume = "89",
pages = "917--930",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "AMER SOC MICROBIOLOGY",
number = "2",

}

RIS (suitable for import to EndNote) - Download

TY - JOUR

T1 - Interactome analysis of the human respiratory syncytial virus RNA polymerase complex identifies protein chaperones as important co-factors that promote L protein stability and RNA synthesis

AU - Munday, Diane Carolyn

AU - Wu, Weining

AU - Smith, Nikki

AU - Fix, Jenna

AU - Noton, Sarah Louise

AU - Galloux, Marie

AU - Touzelet, Olivier

AU - Armstrong, Stuart D

AU - Dawson, Jenna M

AU - Aljabr, Waleed

AU - Easton, Andrew J

AU - Rameix-Welti, Marie-Anne

AU - de Oliveira, Andressa Peres

AU - Simabuco, Fernando

AU - Ventura, Armando M

AU - Hughes, David J

AU - Barr, John N

AU - Fearns, Rachel

AU - Digard, Paul

AU - Eléouët, Jean-François

AU - Hiscox, Julian A

N1 - This research was supported by the award of a Medical Research Council (MRC) Project Grant (MR/K000276/1) to P.D., J.N.B., and J.A.H., an MRC studentship to J.N.B. and J.A.H., NIHR funding to J.A.H., and NIH grant R01AI074903 to R.F. W.A. is supported by the Ministry of Education, Kingdom of Saudi Arabia.

PY - 2015/1/15

Y1 - 2015/1/15

N2 - The human respiratory syncytial virus (HRSV) core viral RNA polymerase comprises the large polymerase protein (L) and its co-factor the phosphoprotein (P) which associate with the viral ribonucleoprotein complex to replicate the genome and, together with the M2-1 protein, transcribe viral mRNAs. Whilst cellular proteins have long been proposed to be involved in the synthesis of HRSV RNA by associating with the polymerase complex, their characterization has been hindered by the difficulty of purifying the viral polymerase from mammalian cell culture. In this study, EGFP-tagged L and P protein expression was coupled with high affinity anti-GFP antibody-based immunoprecipitation and quantitative proteomics to identify cellular proteins that interacted with either the L or the P proteins when expressed as part of a biologically active viral RNP. Several core groups of cellular proteins were identified that interacted with each viral protein, including in both cases, protein chaperones. Ablation of chaperone activity using small molecule inhibitors confirmed previous studies, which suggested this class of proteins acted as positive viral factors. Inhibition of HSP90 chaperone function in the current study showed that HSP90 was critical for L protein function and stability, whether in the presence or absence of the P protein. Inhibition studies suggested that HSP70 also disrupted virus biology and might help the polymerase remodel the nucleocapsid to allow RNA synthesis to occur efficiently. This indicated a pro-viral role for protein chaperones in HRSV replication and demonstrates that the function of cellular proteins can be targeted as potential therapeutics to disrupt virus replication.

AB - The human respiratory syncytial virus (HRSV) core viral RNA polymerase comprises the large polymerase protein (L) and its co-factor the phosphoprotein (P) which associate with the viral ribonucleoprotein complex to replicate the genome and, together with the M2-1 protein, transcribe viral mRNAs. Whilst cellular proteins have long been proposed to be involved in the synthesis of HRSV RNA by associating with the polymerase complex, their characterization has been hindered by the difficulty of purifying the viral polymerase from mammalian cell culture. In this study, EGFP-tagged L and P protein expression was coupled with high affinity anti-GFP antibody-based immunoprecipitation and quantitative proteomics to identify cellular proteins that interacted with either the L or the P proteins when expressed as part of a biologically active viral RNP. Several core groups of cellular proteins were identified that interacted with each viral protein, including in both cases, protein chaperones. Ablation of chaperone activity using small molecule inhibitors confirmed previous studies, which suggested this class of proteins acted as positive viral factors. Inhibition of HSP90 chaperone function in the current study showed that HSP90 was critical for L protein function and stability, whether in the presence or absence of the P protein. Inhibition studies suggested that HSP70 also disrupted virus biology and might help the polymerase remodel the nucleocapsid to allow RNA synthesis to occur efficiently. This indicated a pro-viral role for protein chaperones in HRSV replication and demonstrates that the function of cellular proteins can be targeted as potential therapeutics to disrupt virus replication.

UR - http://www.ncbi.nlm.nih.gov/pubmed/25355874

U2 - 10.1128/JVI.01783-14

DO - 10.1128/JVI.01783-14

M3 - Article

C2 - 25355874

VL - 89

SP - 917

EP - 930

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 2

ER -

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