+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Studying the dynamics of coronavirus replicative structures

Studying the dynamics of coronavirus replicative structures

Methods in Molecular Biology 1282: 261-269

Coronaviruses (CoVs) generate specialized membrane compartments, which consist of double membrane vesicles connected to convoluted membranes, the so-called replicative structures, where viral RNA synthesis takes place. These sites harbor the CoV replication-transcription complexes (RTCs): multi-protein complexes consisting of 16 nonstructural proteins (nsps), the CoV nucleocapsid protein (N) and presumably host proteins. To successfully establish functional membrane-bound RTCs all of the viral and host constituents need to be correctly spatiotemporally organized during viral infection. Few studies, however, have investigated the dynamic processes involved in the formation and functioning of the (subunits of) CoV RTCs and the replicative structures in living cells. In this chapter we describe several protocols to perform time-lapse imaging of CoV-infected cells and to study the kinetics of (subunits of) the CoV replicative structures. The approaches described are not limited to CoV-infected cells; they can also be applied to other virus-infected or non-infected cells.

Please choose payment method:

(PDF emailed within 1 workday: $29.90)

Accession: 058923634

Download citation: RISBibTeXText

PMID: 25720487

Related references

Biogenesis and dynamics of the coronavirus replicative structures. Viruses 4(11): 3245-3269, 2012

Human coronavirus EMC does not require the SARS-coronavirus receptor and maintains broad replicative capability in mammalian cell lines. Mbio 3(6):, 2012

Coronavirus replicative proteins. Nidoviruses): 65-81, 2008

Does form meet function in the coronavirus replicative organelle?. Trends in Microbiology 22(11): 642-647, 2014

Replicative Capacity of MERS Coronavirus in Livestock Cell Lines. Emerging Infectious Diseases 20(2): 276-279, 2014

Coronavirus Nsp10, a critical co-factor for activation of multiple replicative enzymes. Journal of Biological Chemistry 289(37): 25783-25796, 2014

Bioinformatics and functional analyses of coronavirus nonstructural proteins involved in the formation of replicative organelles. Antiviral Research 135: 97, 2016

Coronavirus transcription: subgenomic mouse hepatitis virus replicative intermediates function in RNA synthesis. Journal of Virology 64(3): 1050-1056, 1990

Viscosimetric analysis of the dynamics of alkaline lysis of mammalian cell nuclei as a method of studying radiation damage to DNA structures. Radiobiologiia 24(6): 798-801, 1984

Nascent synthesis of leader sequence-containing subgenomic mRNAs in coronavirus genome-length replicative intermediate RNA. Virology 275(2): 238-243, 2000

Studying the replicative life span of yeast cells. Methods in Molecular Biology 1048: 49-63, 2013

The severe acute respiratory syndrome-coronavirus replicative protein nsp9 is a single-stranded RNA-binding subunit unique in the RNA virus world. Proceedings of the National Academy of Sciences of the United States of America 101(11): 3792-3796, 2004

Chemical evidence for the exclusive occurrence of pppGp nucleoside tetraphosphate in the alkaline hydrolysates of MS2 replicative form and replicative intermediate: implications for the structure of the replicative intermediates and for the mechanism of viral RNA replication. Archives Internationales de Physiologie et de Biochimie 76(5): 963-965, 1968

Studying coronavirus-host protein interactions. Methods in Molecular Biology 1282: 197-212, 2015

Antigenic relationships among proteins of bovine coronavirus, human respiratory coronavirus OC43, and mouse hepatitis coronavirus A59. Journal of Virology 51(2): 384-388, 1984