The Respiratory Syncytial Virus Matrix Protein Possesses 1 a Crm1-Mediated Nuclear Export Mechanism

Reena Ghildyal, Adeline Ho, Manisha Dias, Lydia Soegiyono, Philip Bardin, Kim Tran, Michael Teng, David Jans

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

The respiratory syncytial virus (RSV) matrix (M) protein is localized in the nucleus of infected cells early in infection but is mostly cytoplasmic late in infection. We have previously shown that M localizes in the nucleus through the action of the importin β1 nuclear import receptor. Here, we establish for the first time that M's ability to shuttle to the cytoplasm is due to the action of the nuclear export receptor Crm1, as shown in infected cells, and in cells transfected to express green fluorescent protein (GFP)-M fusion proteins. Specific inhibition of Crm1-mediated nuclear export by leptomycin B increased M nuclear accumulation. Analysis of truncated and point-mutated M derivatives indicated that Crm1-dependent nuclear export of M is attributable to a nuclear export signal (NES) within residues 194 to 206. Importantly, inhibition of M nuclear export resulted in reduced virus production, and a recombinant RSV carrying a mutated NES could not be rescued by reverse genetics. That this is likely to be due to the inability of a nuclear export deficient M to localize to regions of virus assembly is indicated by the fact that a nuclear-export-deficient GFP-M fails to localize to regions of virus assembly when expressed in cells infected with wild-type RSV. Together, our data suggest that Crm1-dependent nuclear export of M is central to RSV infection, representing the first report of such a mechanism for a paramyxovirus M protein and with important implications for related paramyxoviruses.
Original languageEnglish
Pages (from-to)5353-5362
Number of pages10
JournalJournal of Virology
Volume83
Issue number11
DOIs
Publication statusPublished - 2009

Fingerprint

Respiratory Syncytial Viruses
Cell Nucleus Active Transport
viruses
Proteins
proteins
Nuclear Export Signals
Virus Assembly
Respirovirus
Cytoplasmic and Nuclear Receptors
Green Fluorescent Proteins
green fluorescent protein
Karyopherins
importins
Reverse Genetics
Respiratory Syncytial Virus Infections
infection
receptors
Infection
Cell Nucleus
cells

Cite this

Ghildyal, Reena ; Ho, Adeline ; Dias, Manisha ; Soegiyono, Lydia ; Bardin, Philip ; Tran, Kim ; Teng, Michael ; Jans, David. / The Respiratory Syncytial Virus Matrix Protein Possesses 1 a Crm1-Mediated Nuclear Export Mechanism. In: Journal of Virology. 2009 ; Vol. 83, No. 11. pp. 5353-5362.
@article{67cbf0dbce3d4c52a381340d090563bc,
title = "The Respiratory Syncytial Virus Matrix Protein Possesses 1 a Crm1-Mediated Nuclear Export Mechanism",
abstract = "The respiratory syncytial virus (RSV) matrix (M) protein is localized in the nucleus of infected cells early in infection but is mostly cytoplasmic late in infection. We have previously shown that M localizes in the nucleus through the action of the importin β1 nuclear import receptor. Here, we establish for the first time that M's ability to shuttle to the cytoplasm is due to the action of the nuclear export receptor Crm1, as shown in infected cells, and in cells transfected to express green fluorescent protein (GFP)-M fusion proteins. Specific inhibition of Crm1-mediated nuclear export by leptomycin B increased M nuclear accumulation. Analysis of truncated and point-mutated M derivatives indicated that Crm1-dependent nuclear export of M is attributable to a nuclear export signal (NES) within residues 194 to 206. Importantly, inhibition of M nuclear export resulted in reduced virus production, and a recombinant RSV carrying a mutated NES could not be rescued by reverse genetics. That this is likely to be due to the inability of a nuclear export deficient M to localize to regions of virus assembly is indicated by the fact that a nuclear-export-deficient GFP-M fails to localize to regions of virus assembly when expressed in cells infected with wild-type RSV. Together, our data suggest that Crm1-dependent nuclear export of M is central to RSV infection, representing the first report of such a mechanism for a paramyxovirus M protein and with important implications for related paramyxoviruses.",
author = "Reena Ghildyal and Adeline Ho and Manisha Dias and Lydia Soegiyono and Philip Bardin and Kim Tran and Michael Teng and David Jans",
year = "2009",
doi = "10.1128/JVI.02374-08",
language = "English",
volume = "83",
pages = "5353--5362",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "11",

}

Ghildyal, R, Ho, A, Dias, M, Soegiyono, L, Bardin, P, Tran, K, Teng, M & Jans, D 2009, 'The Respiratory Syncytial Virus Matrix Protein Possesses 1 a Crm1-Mediated Nuclear Export Mechanism', Journal of Virology, vol. 83, no. 11, pp. 5353-5362. https://doi.org/10.1128/JVI.02374-08

The Respiratory Syncytial Virus Matrix Protein Possesses 1 a Crm1-Mediated Nuclear Export Mechanism. / Ghildyal, Reena; Ho, Adeline; Dias, Manisha; Soegiyono, Lydia; Bardin, Philip; Tran, Kim; Teng, Michael; Jans, David.

In: Journal of Virology, Vol. 83, No. 11, 2009, p. 5353-5362.

Research output: Contribution to journalArticle

TY - JOUR

T1 - The Respiratory Syncytial Virus Matrix Protein Possesses 1 a Crm1-Mediated Nuclear Export Mechanism

AU - Ghildyal, Reena

AU - Ho, Adeline

AU - Dias, Manisha

AU - Soegiyono, Lydia

AU - Bardin, Philip

AU - Tran, Kim

AU - Teng, Michael

AU - Jans, David

PY - 2009

Y1 - 2009

N2 - The respiratory syncytial virus (RSV) matrix (M) protein is localized in the nucleus of infected cells early in infection but is mostly cytoplasmic late in infection. We have previously shown that M localizes in the nucleus through the action of the importin β1 nuclear import receptor. Here, we establish for the first time that M's ability to shuttle to the cytoplasm is due to the action of the nuclear export receptor Crm1, as shown in infected cells, and in cells transfected to express green fluorescent protein (GFP)-M fusion proteins. Specific inhibition of Crm1-mediated nuclear export by leptomycin B increased M nuclear accumulation. Analysis of truncated and point-mutated M derivatives indicated that Crm1-dependent nuclear export of M is attributable to a nuclear export signal (NES) within residues 194 to 206. Importantly, inhibition of M nuclear export resulted in reduced virus production, and a recombinant RSV carrying a mutated NES could not be rescued by reverse genetics. That this is likely to be due to the inability of a nuclear export deficient M to localize to regions of virus assembly is indicated by the fact that a nuclear-export-deficient GFP-M fails to localize to regions of virus assembly when expressed in cells infected with wild-type RSV. Together, our data suggest that Crm1-dependent nuclear export of M is central to RSV infection, representing the first report of such a mechanism for a paramyxovirus M protein and with important implications for related paramyxoviruses.

AB - The respiratory syncytial virus (RSV) matrix (M) protein is localized in the nucleus of infected cells early in infection but is mostly cytoplasmic late in infection. We have previously shown that M localizes in the nucleus through the action of the importin β1 nuclear import receptor. Here, we establish for the first time that M's ability to shuttle to the cytoplasm is due to the action of the nuclear export receptor Crm1, as shown in infected cells, and in cells transfected to express green fluorescent protein (GFP)-M fusion proteins. Specific inhibition of Crm1-mediated nuclear export by leptomycin B increased M nuclear accumulation. Analysis of truncated and point-mutated M derivatives indicated that Crm1-dependent nuclear export of M is attributable to a nuclear export signal (NES) within residues 194 to 206. Importantly, inhibition of M nuclear export resulted in reduced virus production, and a recombinant RSV carrying a mutated NES could not be rescued by reverse genetics. That this is likely to be due to the inability of a nuclear export deficient M to localize to regions of virus assembly is indicated by the fact that a nuclear-export-deficient GFP-M fails to localize to regions of virus assembly when expressed in cells infected with wild-type RSV. Together, our data suggest that Crm1-dependent nuclear export of M is central to RSV infection, representing the first report of such a mechanism for a paramyxovirus M protein and with important implications for related paramyxoviruses.

U2 - 10.1128/JVI.02374-08

DO - 10.1128/JVI.02374-08

M3 - Article

VL - 83

SP - 5353

EP - 5362

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 11

ER -