NC-mediated nucleolar localization of retroviral gag proteins.

Timothy L. Lochmann, Darrin V. Bann, Eileen P. Ryan, Andrea R. Beyer, Annie Mao, Alan Cochrane, Leslie Parent

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

The assembly and release of retrovirus particles from the cell membrane is directed by the Gag polyprotein. The Gag protein of Rous sarcoma virus (RSV) traffics through the nucleus prior to plasma membrane localization. We previously reported that nuclear localization of RSV Gag is linked to efficient packaging of viral genomic RNA, however the intranuclear activities of RSV Gag are not well understood. To gain insight into the properties of the RSV Gag protein within the nucleus, we examined the subnuclear localization and dynamic trafficking of RSV Gag. Restriction of RSV Gag to the nucleus by mutating its nuclear export signal (NES) in the p10 domain or interfering with CRM1-mediated nuclear export of Gag by leptomycin B (LMB) treatment led to the accumulation of Gag in nucleoli and discrete nucleoplasmic foci. Retention of RSV Gag in nucleoli was reduced with cis-expression of the 5' untranslated RU5 region of the viral RNA genome, suggesting the psi (Ψ) packaging signal may alter the subnuclear localization of Gag. Fluorescence recovery after photobleaching (FRAP) demonstrated that the nucleolar fraction of Gag was highly mobile, indicating that there was rapid exchange with Gag proteins in the nucleoplasm. RSV Gag is targeted to nucleoli by a nucleolar localization signal (NoLS) in the NC domain, and similarly, the human immunodeficiency virus type 1 (HIV-1) NC protein also contains an NoLS consisting of basic residues. Interestingly, co-expression of HIV-1 NC or Rev with HIV-1 Gag resulted in accumulation of Gag in nucleoli. Moreover, a subpopulation of HIV-1 Gag was detected in the nucleoli of HeLa cells stably expressing the entire HIV-1 genome in a Rev-dependent fashion. These findings suggest that the RSV and HIV-1 Gag proteins undergo nucleolar trafficking in the setting of viral infection.

Original languageEnglish (US)
Pages (from-to)304-318
Number of pages15
JournalUnknown Journal
Volume171
Issue number2
StatePublished - Jan 1 2013

Fingerprint

gag Gene Products
Rous sarcoma virus
HIV-1
Viral RNA
Product Packaging
Human Immunodeficiency Virus gag Gene Products
Cell Membrane
Nuclear Export Signals
Fluorescence Recovery After Photobleaching
Cell Nucleus Active Transport
Viral Genome
5' Untranslated Regions
Virus Diseases
Retroviridae
HeLa Cells
Genome

All Science Journal Classification (ASJC) codes

  • Virology
  • Infectious Diseases
  • Cancer Research

Cite this

Lochmann, T. L., Bann, D. V., Ryan, E. P., Beyer, A. R., Mao, A., Cochrane, A., & Parent, L. (2013). NC-mediated nucleolar localization of retroviral gag proteins. Unknown Journal, 171(2), 304-318.
Lochmann, Timothy L. ; Bann, Darrin V. ; Ryan, Eileen P. ; Beyer, Andrea R. ; Mao, Annie ; Cochrane, Alan ; Parent, Leslie. / NC-mediated nucleolar localization of retroviral gag proteins. In: Unknown Journal. 2013 ; Vol. 171, No. 2. pp. 304-318.
@article{bc59e5fb5a8c470e9ff6d7be7775053c,
title = "NC-mediated nucleolar localization of retroviral gag proteins.",
abstract = "The assembly and release of retrovirus particles from the cell membrane is directed by the Gag polyprotein. The Gag protein of Rous sarcoma virus (RSV) traffics through the nucleus prior to plasma membrane localization. We previously reported that nuclear localization of RSV Gag is linked to efficient packaging of viral genomic RNA, however the intranuclear activities of RSV Gag are not well understood. To gain insight into the properties of the RSV Gag protein within the nucleus, we examined the subnuclear localization and dynamic trafficking of RSV Gag. Restriction of RSV Gag to the nucleus by mutating its nuclear export signal (NES) in the p10 domain or interfering with CRM1-mediated nuclear export of Gag by leptomycin B (LMB) treatment led to the accumulation of Gag in nucleoli and discrete nucleoplasmic foci. Retention of RSV Gag in nucleoli was reduced with cis-expression of the 5' untranslated RU5 region of the viral RNA genome, suggesting the psi (Ψ) packaging signal may alter the subnuclear localization of Gag. Fluorescence recovery after photobleaching (FRAP) demonstrated that the nucleolar fraction of Gag was highly mobile, indicating that there was rapid exchange with Gag proteins in the nucleoplasm. RSV Gag is targeted to nucleoli by a nucleolar localization signal (NoLS) in the NC domain, and similarly, the human immunodeficiency virus type 1 (HIV-1) NC protein also contains an NoLS consisting of basic residues. Interestingly, co-expression of HIV-1 NC or Rev with HIV-1 Gag resulted in accumulation of Gag in nucleoli. Moreover, a subpopulation of HIV-1 Gag was detected in the nucleoli of HeLa cells stably expressing the entire HIV-1 genome in a Rev-dependent fashion. These findings suggest that the RSV and HIV-1 Gag proteins undergo nucleolar trafficking in the setting of viral infection.",
author = "Lochmann, {Timothy L.} and Bann, {Darrin V.} and Ryan, {Eileen P.} and Beyer, {Andrea R.} and Annie Mao and Alan Cochrane and Leslie Parent",
year = "2013",
month = "1",
day = "1",
language = "English (US)",
volume = "171",
pages = "304--318",
journal = "[No source information available]",
issn = "0042-1215",
number = "2",

}

Lochmann, TL, Bann, DV, Ryan, EP, Beyer, AR, Mao, A, Cochrane, A & Parent, L 2013, 'NC-mediated nucleolar localization of retroviral gag proteins.', Unknown Journal, vol. 171, no. 2, pp. 304-318.

NC-mediated nucleolar localization of retroviral gag proteins. / Lochmann, Timothy L.; Bann, Darrin V.; Ryan, Eileen P.; Beyer, Andrea R.; Mao, Annie; Cochrane, Alan; Parent, Leslie.

In: Unknown Journal, Vol. 171, No. 2, 01.01.2013, p. 304-318.

Research output: Contribution to journalArticle

TY - JOUR

T1 - NC-mediated nucleolar localization of retroviral gag proteins.

AU - Lochmann, Timothy L.

AU - Bann, Darrin V.

AU - Ryan, Eileen P.

AU - Beyer, Andrea R.

AU - Mao, Annie

AU - Cochrane, Alan

AU - Parent, Leslie

PY - 2013/1/1

Y1 - 2013/1/1

N2 - The assembly and release of retrovirus particles from the cell membrane is directed by the Gag polyprotein. The Gag protein of Rous sarcoma virus (RSV) traffics through the nucleus prior to plasma membrane localization. We previously reported that nuclear localization of RSV Gag is linked to efficient packaging of viral genomic RNA, however the intranuclear activities of RSV Gag are not well understood. To gain insight into the properties of the RSV Gag protein within the nucleus, we examined the subnuclear localization and dynamic trafficking of RSV Gag. Restriction of RSV Gag to the nucleus by mutating its nuclear export signal (NES) in the p10 domain or interfering with CRM1-mediated nuclear export of Gag by leptomycin B (LMB) treatment led to the accumulation of Gag in nucleoli and discrete nucleoplasmic foci. Retention of RSV Gag in nucleoli was reduced with cis-expression of the 5' untranslated RU5 region of the viral RNA genome, suggesting the psi (Ψ) packaging signal may alter the subnuclear localization of Gag. Fluorescence recovery after photobleaching (FRAP) demonstrated that the nucleolar fraction of Gag was highly mobile, indicating that there was rapid exchange with Gag proteins in the nucleoplasm. RSV Gag is targeted to nucleoli by a nucleolar localization signal (NoLS) in the NC domain, and similarly, the human immunodeficiency virus type 1 (HIV-1) NC protein also contains an NoLS consisting of basic residues. Interestingly, co-expression of HIV-1 NC or Rev with HIV-1 Gag resulted in accumulation of Gag in nucleoli. Moreover, a subpopulation of HIV-1 Gag was detected in the nucleoli of HeLa cells stably expressing the entire HIV-1 genome in a Rev-dependent fashion. These findings suggest that the RSV and HIV-1 Gag proteins undergo nucleolar trafficking in the setting of viral infection.

AB - The assembly and release of retrovirus particles from the cell membrane is directed by the Gag polyprotein. The Gag protein of Rous sarcoma virus (RSV) traffics through the nucleus prior to plasma membrane localization. We previously reported that nuclear localization of RSV Gag is linked to efficient packaging of viral genomic RNA, however the intranuclear activities of RSV Gag are not well understood. To gain insight into the properties of the RSV Gag protein within the nucleus, we examined the subnuclear localization and dynamic trafficking of RSV Gag. Restriction of RSV Gag to the nucleus by mutating its nuclear export signal (NES) in the p10 domain or interfering with CRM1-mediated nuclear export of Gag by leptomycin B (LMB) treatment led to the accumulation of Gag in nucleoli and discrete nucleoplasmic foci. Retention of RSV Gag in nucleoli was reduced with cis-expression of the 5' untranslated RU5 region of the viral RNA genome, suggesting the psi (Ψ) packaging signal may alter the subnuclear localization of Gag. Fluorescence recovery after photobleaching (FRAP) demonstrated that the nucleolar fraction of Gag was highly mobile, indicating that there was rapid exchange with Gag proteins in the nucleoplasm. RSV Gag is targeted to nucleoli by a nucleolar localization signal (NoLS) in the NC domain, and similarly, the human immunodeficiency virus type 1 (HIV-1) NC protein also contains an NoLS consisting of basic residues. Interestingly, co-expression of HIV-1 NC or Rev with HIV-1 Gag resulted in accumulation of Gag in nucleoli. Moreover, a subpopulation of HIV-1 Gag was detected in the nucleoli of HeLa cells stably expressing the entire HIV-1 genome in a Rev-dependent fashion. These findings suggest that the RSV and HIV-1 Gag proteins undergo nucleolar trafficking in the setting of viral infection.

UR - http://www.scopus.com/inward/record.url?scp=85027936717&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85027936717&partnerID=8YFLogxK

M3 - Article

C2 - 23036987

AN - SCOPUS:85027936717

VL - 171

SP - 304

EP - 318

JO - [No source information available]

JF - [No source information available]

SN - 0042-1215

IS - 2

ER -

Lochmann TL, Bann DV, Ryan EP, Beyer AR, Mao A, Cochrane A et al. NC-mediated nucleolar localization of retroviral gag proteins. Unknown Journal. 2013 Jan 1;171(2):304-318.