Studying the dynamics of coronavirus replicative structures

Marne C. Hagemeijer*, Cornelis A M De Haan

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

Abstract

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.

Original languageEnglish
Title of host publicationCoronaviruses: Methods and Protocols
Editors Helena Jane Maier, Erica Bickerton, Paul Britton
PublisherSpringer New York
Pages261-269
Number of pages9
VolumeV
ISBN (Electronic)9781493924387
ISBN (Print)9781493924370
DOIs
Publication statusPublished - 26 Feb 2015

Publication series

NameMethods in Molecular Biology
Volume1282
ISSN (Print)1064-3745

Keywords

  • Coronavirus
  • Dynamics
  • Fluorescence loss in photobleaching
  • Fluorescence recovery after photobleaching
  • Live-cell imaging
  • Nonstructural proteins
  • Replication-transcription complex

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