Dynamics of coronavirus replication-transcription complexes

Autor: Hagemeijer, Marne C, Verheije, Monique H, Ulasli, Mustafa, Shaltiël, Indra A, de Vries, Lisa A, Reggiori, Fulvio, Rottier, Peter J M, de Haan, Cornelis A M, LS Pathologie, LS Virologie
Přispěvatelé: LS Pathologie, LS Virologie
Jazyk: angličtina
Rok vydání: 2010
Předmět:
Cytoplasmic Vesicles/metabolism
Transcription
Genetic
viruses
Murine hepatitis virus/genetics
Viral Nonstructural Proteins
medicine.disease_cause
Virus Replication
Green Fluorescent Proteins/genetics
Mice
Transcription (biology)
Coronaviridae
Viral
Coronavirus
Microscopy
medicine.diagnostic_test
virus diseases
Recombinant Fusion Proteins/chemistry
Genome Replication and Regulation of Viral Gene Expression
Cell biology
Viral Nonstructural Proteins/chemistry
Host-Pathogen Interactions
Virus Replication/genetics
Coronavirus Infections/metabolism
Coronavirus Infections
Transcription
Macromolecular Substances
Recombinant Fusion Proteins
Immunology
Green Fluorescent Proteins
Biology
Immunofluorescence
Electron
Microbiology
Virus
Cell Line
Genetic
Microscopy
Electron
Transmission
Virology
medicine
Transmission
Animals
Humans
DNA Primers
DNA Primers/genetics
Murine hepatitis virus
Base Sequence
Cytoplasmic Vesicles
RNA
DNA
DNA
Viral/genetics
biology.organism_classification
Viral replication
Cytoplasm
Insect Science
DNA
Viral
Cats
HeLa Cells
Zdroj: Hagemeijer, M C, Verheije, M H, Ulasli, M, Shaltiël, I A, de Vries, L A, Reggiori, F, Rottier, P J M & de Haan, C A M 2010, ' Dynamics of coronavirus replication-transcription complexes ', Journal of Virology, vol. 84, no. 4, pp. 2134-49 . https://doi.org/10.1128/JVI.01716-09
Journal of Virology
Journal of Virology, 84(4), 2134. American Society for Microbiology
ISSN: 0022-538X
DOI: 10.1128/JVI.01716-09
Popis: Coronaviruses induce in infected cells the formation of double-membrane vesicles (DMVs) in which the replication-transcription complexes (RTCs) are anchored. To study the dynamics of these coronavirus replicative structures, we generated recombinant murine hepatitis coronaviruses that express tagged versions of the nonstructural protein nsp2. We demonstrated by using immunofluorescence assays and electron microscopy that this protein is recruited to the DMV-anchored RTCs, for which its C terminus is essential. Live-cell imaging of infected cells demonstrated that small nsp2-positive structures move through the cytoplasm in a microtubule-dependent manner. In contrast, large fluorescent structures are rather immobile. Microtubule-mediated transport of DMVs, however, is not required for efficient replication. Biochemical analyses indicated that the nsp2 protein is associated with the cytoplasmic side of the DMVs. Yet, no recovery of fluorescence was observed when (part of) the nsp2-positive foci were bleached. This result was confirmed by the observation that preexisting RTCs did not exchange fluorescence after fusion of cells expressing either a green or a red fluorescent nsp2. Apparently, nsp2, once recruited to the RTCs, is not exchanged with nsp2 present in the cytoplasm or at other DMVs. Our data show a remarkable resemblance to results obtained recently by others with hepatitis C virus. The observations point to intriguing and as yet unrecognized similarities between the RTC dynamics of different plus-strand RNA viruses.
Databáze: OpenAIRE
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