Movements of HIV-1 genomic RNA-APOBEC3F complexes and PKR reveal cytoplasmic and nuclear PKR defenses and HIV-1 evasion strategies

Mariana Marin, Sheetal Golem, Susan L. Kozak, David Kabat

Research output: Contribution to journalArticle

Abstract

APOBEC3 cytidine deaminases and viral genomic RNA (gRNA) occur in virions, polysomes, and cytoplasmic granules, but have not been tracked together. Moreover, gRNA traffic is important, but the factors that move it into granules are unknown. Using in situ hybridization of transfected cells and protein synthesis inhibitors that drive mRNAs between locales, we observed APOBEC3F cotrafficking with gRNA without altering its movements. Whereas cells with little cytoplasmic gRNA were translationally active and accumulated Gag, suprathreshold amounts induced autophosphorylation of the cytoplasmic double-stranded RNA (dsRNA)-dependent protein kinase (PKR), causing eIF2α phosphorylation, protein synthesis suppression, and gRNA sequestration in stress granules. Additionally, we confirmed recent evidence that PKR is activated by chromosome-associated cellular dsRNAs after nuclear membranes disperse in prophase. By arresting cells in G2, HIV-1 blocks this mechanism for PKR activation and eIF2α phosphorylation. However, cytopathic membrane damage in CD4- and coreceptor-positive cultures infected with laboratory-adapted fusogenic HIV-1LAI eventually enabled PKR entry and activation in interphase nuclei. These results reveal multiple stages in the PKR-HIV-1 battleground that culminate in cell death. We discuss evidence suggesting that HIV-1s evolve in vivo to prevent or delay PKR activation by all these mechanisms.

Original languageEnglish (US)
Pages (from-to)124-139
Number of pages16
JournalVirus Research
Volume213
DOIs
StatePublished - Feb 2 2016

Fingerprint

HIV-1
RNA
eIF-2 Kinase
Phosphorylation
HIV
Cytidine Deaminase
Cytoplasmic Granules
Prophase
Protein Synthesis Inhibitors
Polyribosomes
Double-Stranded RNA
Interphase
Nuclear Envelope
Viral RNA
Virion
In Situ Hybridization
Cell Death
Chromosomes
Messenger RNA
Membranes

Keywords

  • APOBEC3F
  • HIV-1
  • Mitosis
  • Nuclear pores
  • P-bodies
  • PKR
  • Stress granules

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases
  • Cancer Research

Cite this

Movements of HIV-1 genomic RNA-APOBEC3F complexes and PKR reveal cytoplasmic and nuclear PKR defenses and HIV-1 evasion strategies. / Marin, Mariana; Golem, Sheetal; Kozak, Susan L.; Kabat, David.

In: Virus Research, Vol. 213, 02.02.2016, p. 124-139.

Research output: Contribution to journalArticle

Marin, Mariana ; Golem, Sheetal ; Kozak, Susan L. ; Kabat, David. / Movements of HIV-1 genomic RNA-APOBEC3F complexes and PKR reveal cytoplasmic and nuclear PKR defenses and HIV-1 evasion strategies. In: Virus Research. 2016 ; Vol. 213. pp. 124-139.
@article{c140e77941d94e75a4c6f2c39b5926e9,
title = "Movements of HIV-1 genomic RNA-APOBEC3F complexes and PKR reveal cytoplasmic and nuclear PKR defenses and HIV-1 evasion strategies",
abstract = "APOBEC3 cytidine deaminases and viral genomic RNA (gRNA) occur in virions, polysomes, and cytoplasmic granules, but have not been tracked together. Moreover, gRNA traffic is important, but the factors that move it into granules are unknown. Using in situ hybridization of transfected cells and protein synthesis inhibitors that drive mRNAs between locales, we observed APOBEC3F cotrafficking with gRNA without altering its movements. Whereas cells with little cytoplasmic gRNA were translationally active and accumulated Gag, suprathreshold amounts induced autophosphorylation of the cytoplasmic double-stranded RNA (dsRNA)-dependent protein kinase (PKR), causing eIF2α phosphorylation, protein synthesis suppression, and gRNA sequestration in stress granules. Additionally, we confirmed recent evidence that PKR is activated by chromosome-associated cellular dsRNAs after nuclear membranes disperse in prophase. By arresting cells in G2, HIV-1 blocks this mechanism for PKR activation and eIF2α phosphorylation. However, cytopathic membrane damage in CD4- and coreceptor-positive cultures infected with laboratory-adapted fusogenic HIV-1LAI eventually enabled PKR entry and activation in interphase nuclei. These results reveal multiple stages in the PKR-HIV-1 battleground that culminate in cell death. We discuss evidence suggesting that HIV-1s evolve in vivo to prevent or delay PKR activation by all these mechanisms.",
keywords = "APOBEC3F, HIV-1, Mitosis, Nuclear pores, P-bodies, PKR, Stress granules",
author = "Mariana Marin and Sheetal Golem and Kozak, {Susan L.} and David Kabat",
year = "2016",
month = "2",
day = "2",
doi = "10.1016/j.virusres.2015.11.023",
language = "English (US)",
volume = "213",
pages = "124--139",
journal = "Virus Research",
issn = "0168-1702",
publisher = "Elsevier",

}

TY - JOUR

T1 - Movements of HIV-1 genomic RNA-APOBEC3F complexes and PKR reveal cytoplasmic and nuclear PKR defenses and HIV-1 evasion strategies

AU - Marin, Mariana

AU - Golem, Sheetal

AU - Kozak, Susan L.

AU - Kabat, David

PY - 2016/2/2

Y1 - 2016/2/2

N2 - APOBEC3 cytidine deaminases and viral genomic RNA (gRNA) occur in virions, polysomes, and cytoplasmic granules, but have not been tracked together. Moreover, gRNA traffic is important, but the factors that move it into granules are unknown. Using in situ hybridization of transfected cells and protein synthesis inhibitors that drive mRNAs between locales, we observed APOBEC3F cotrafficking with gRNA without altering its movements. Whereas cells with little cytoplasmic gRNA were translationally active and accumulated Gag, suprathreshold amounts induced autophosphorylation of the cytoplasmic double-stranded RNA (dsRNA)-dependent protein kinase (PKR), causing eIF2α phosphorylation, protein synthesis suppression, and gRNA sequestration in stress granules. Additionally, we confirmed recent evidence that PKR is activated by chromosome-associated cellular dsRNAs after nuclear membranes disperse in prophase. By arresting cells in G2, HIV-1 blocks this mechanism for PKR activation and eIF2α phosphorylation. However, cytopathic membrane damage in CD4- and coreceptor-positive cultures infected with laboratory-adapted fusogenic HIV-1LAI eventually enabled PKR entry and activation in interphase nuclei. These results reveal multiple stages in the PKR-HIV-1 battleground that culminate in cell death. We discuss evidence suggesting that HIV-1s evolve in vivo to prevent or delay PKR activation by all these mechanisms.

AB - APOBEC3 cytidine deaminases and viral genomic RNA (gRNA) occur in virions, polysomes, and cytoplasmic granules, but have not been tracked together. Moreover, gRNA traffic is important, but the factors that move it into granules are unknown. Using in situ hybridization of transfected cells and protein synthesis inhibitors that drive mRNAs between locales, we observed APOBEC3F cotrafficking with gRNA without altering its movements. Whereas cells with little cytoplasmic gRNA were translationally active and accumulated Gag, suprathreshold amounts induced autophosphorylation of the cytoplasmic double-stranded RNA (dsRNA)-dependent protein kinase (PKR), causing eIF2α phosphorylation, protein synthesis suppression, and gRNA sequestration in stress granules. Additionally, we confirmed recent evidence that PKR is activated by chromosome-associated cellular dsRNAs after nuclear membranes disperse in prophase. By arresting cells in G2, HIV-1 blocks this mechanism for PKR activation and eIF2α phosphorylation. However, cytopathic membrane damage in CD4- and coreceptor-positive cultures infected with laboratory-adapted fusogenic HIV-1LAI eventually enabled PKR entry and activation in interphase nuclei. These results reveal multiple stages in the PKR-HIV-1 battleground that culminate in cell death. We discuss evidence suggesting that HIV-1s evolve in vivo to prevent or delay PKR activation by all these mechanisms.

KW - APOBEC3F

KW - HIV-1

KW - Mitosis

KW - Nuclear pores

KW - P-bodies

KW - PKR

KW - Stress granules

UR - http://www.scopus.com/inward/record.url?scp=84958769502&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84958769502&partnerID=8YFLogxK

U2 - 10.1016/j.virusres.2015.11.023

DO - 10.1016/j.virusres.2015.11.023

M3 - Article

C2 - 26626364

AN - SCOPUS:84958769502

VL - 213

SP - 124

EP - 139

JO - Virus Research

JF - Virus Research

SN - 0168-1702

ER -