Cargando…

Three-Dimensional Analysis of a Viral RNA Replication Complex Reveals a Virus-Induced Mini-Organelle

Positive-strand RNA viruses are the largest genetic class of viruses and include many serious human pathogens. All positive-strand RNA viruses replicate their genomes in association with intracellular membrane rearrangements such as single- or double-membrane vesicles. However, the exact sites of RN...

Descripción completa

Detalles Bibliográficos
Autores principales: Kopek, Benjamin G, Perkins, Guy, Miller, David J, Ellisman, Mark H, Ahlquist, Paul
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1945040/
https://www.ncbi.nlm.nih.gov/pubmed/17696647
http://dx.doi.org/10.1371/journal.pbio.0050220
_version_ 1782134485324660736
author Kopek, Benjamin G
Perkins, Guy
Miller, David J
Ellisman, Mark H
Ahlquist, Paul
author_facet Kopek, Benjamin G
Perkins, Guy
Miller, David J
Ellisman, Mark H
Ahlquist, Paul
author_sort Kopek, Benjamin G
collection PubMed
description Positive-strand RNA viruses are the largest genetic class of viruses and include many serious human pathogens. All positive-strand RNA viruses replicate their genomes in association with intracellular membrane rearrangements such as single- or double-membrane vesicles. However, the exact sites of RNA synthesis and crucial topological relationships between relevant membranes, vesicle interiors, surrounding lumens, and cytoplasm generally are poorly defined. We applied electron microscope tomography and complementary approaches to flock house virus (FHV)–infected Drosophila cells to provide the first 3-D analysis of such replication complexes. The sole FHV RNA replication factor, protein A, and FHV-specific 5-bromouridine 5'-triphosphate incorporation localized between inner and outer mitochondrial membranes inside ∼50-nm vesicles (spherules), which thus are FHV-induced compartments for viral RNA synthesis. All such FHV spherules were outer mitochondrial membrane invaginations with interiors connected to the cytoplasm by a necked channel of ∼10-nm diameter, which is sufficient for ribonucleotide import and product RNA export. Tomographic, biochemical, and other results imply that FHV spherules contain, on average, three RNA replication intermediates and an interior shell of ∼100 membrane-spanning, self-interacting protein As. The results identify spherules as the site of protein A and nascent RNA accumulation and define spherule topology, dimensions, and stoichiometry to reveal the nature and many details of the organization and function of the FHV RNA replication complex. The resulting insights appear relevant to many other positive-strand RNA viruses and support recently proposed structural and likely evolutionary parallels with retrovirus and double-stranded RNA virus virions.
format Text
id pubmed-1945040
institution National Center for Biotechnology Information
language English
publishDate 2007
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-19450402007-08-14 Three-Dimensional Analysis of a Viral RNA Replication Complex Reveals a Virus-Induced Mini-Organelle Kopek, Benjamin G Perkins, Guy Miller, David J Ellisman, Mark H Ahlquist, Paul PLoS Biol Research Article Positive-strand RNA viruses are the largest genetic class of viruses and include many serious human pathogens. All positive-strand RNA viruses replicate their genomes in association with intracellular membrane rearrangements such as single- or double-membrane vesicles. However, the exact sites of RNA synthesis and crucial topological relationships between relevant membranes, vesicle interiors, surrounding lumens, and cytoplasm generally are poorly defined. We applied electron microscope tomography and complementary approaches to flock house virus (FHV)–infected Drosophila cells to provide the first 3-D analysis of such replication complexes. The sole FHV RNA replication factor, protein A, and FHV-specific 5-bromouridine 5'-triphosphate incorporation localized between inner and outer mitochondrial membranes inside ∼50-nm vesicles (spherules), which thus are FHV-induced compartments for viral RNA synthesis. All such FHV spherules were outer mitochondrial membrane invaginations with interiors connected to the cytoplasm by a necked channel of ∼10-nm diameter, which is sufficient for ribonucleotide import and product RNA export. Tomographic, biochemical, and other results imply that FHV spherules contain, on average, three RNA replication intermediates and an interior shell of ∼100 membrane-spanning, self-interacting protein As. The results identify spherules as the site of protein A and nascent RNA accumulation and define spherule topology, dimensions, and stoichiometry to reveal the nature and many details of the organization and function of the FHV RNA replication complex. The resulting insights appear relevant to many other positive-strand RNA viruses and support recently proposed structural and likely evolutionary parallels with retrovirus and double-stranded RNA virus virions. Public Library of Science 2007-09 2007-08-14 /pmc/articles/PMC1945040/ /pubmed/17696647 http://dx.doi.org/10.1371/journal.pbio.0050220 Text en © 2007 Kopek et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Kopek, Benjamin G
Perkins, Guy
Miller, David J
Ellisman, Mark H
Ahlquist, Paul
Three-Dimensional Analysis of a Viral RNA Replication Complex Reveals a Virus-Induced Mini-Organelle
title Three-Dimensional Analysis of a Viral RNA Replication Complex Reveals a Virus-Induced Mini-Organelle
title_full Three-Dimensional Analysis of a Viral RNA Replication Complex Reveals a Virus-Induced Mini-Organelle
title_fullStr Three-Dimensional Analysis of a Viral RNA Replication Complex Reveals a Virus-Induced Mini-Organelle
title_full_unstemmed Three-Dimensional Analysis of a Viral RNA Replication Complex Reveals a Virus-Induced Mini-Organelle
title_short Three-Dimensional Analysis of a Viral RNA Replication Complex Reveals a Virus-Induced Mini-Organelle
title_sort three-dimensional analysis of a viral rna replication complex reveals a virus-induced mini-organelle
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1945040/
https://www.ncbi.nlm.nih.gov/pubmed/17696647
http://dx.doi.org/10.1371/journal.pbio.0050220
work_keys_str_mv AT kopekbenjaming threedimensionalanalysisofaviralrnareplicationcomplexrevealsavirusinducedminiorganelle
AT perkinsguy threedimensionalanalysisofaviralrnareplicationcomplexrevealsavirusinducedminiorganelle
AT millerdavidj threedimensionalanalysisofaviralrnareplicationcomplexrevealsavirusinducedminiorganelle
AT ellismanmarkh threedimensionalanalysisofaviralrnareplicationcomplexrevealsavirusinducedminiorganelle
AT ahlquistpaul threedimensionalanalysisofaviralrnareplicationcomplexrevealsavirusinducedminiorganelle