RNA replication of mouse hepatitis virus takes place at double-membrane vesicles

Journal of Virology
R GosertS C Baker

Abstract

The replication complexes (RCs) of positive-stranded RNA viruses are intimately associated with cellular membranes. To investigate membrane alterations and to characterize the RC of mouse hepatitis virus (MHV), we performed biochemical and ultrastructural studies using MHV-infected cells. Biochemical fractionation showed that all 10 of the MHV gene 1 polyprotein products examined pelleted with the membrane fraction, consistent with membrane association of the RC. Furthermore, MHV gene 1 products p290, p210, and p150 and the p150 cleavage product membrane protein 1 (MP1, also called p44) were resistant to extraction with Triton X-114, indicating that they are integral membrane proteins. The ultrastructural analysis revealed double-membrane vesicles (DMVs) in the cytoplasm of MHV-infected cells. The DMVs were found either as separate entities or as small clusters of vesicles. To determine whether MHV proteins and viral RNA were associated with the DMVs, we performed immunocytochemistry electron microscopy (IEM). We found that the DMVs were labeled using an antiserum directed against proteins derived from open reading frame 1a of MHV. By electron microscopy in situ hybridization (ISH) using MHV-specific RNA probes, DMVs were highl...Continue Reading

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Citations

Nov 25, 2003·Journal of Biomedical Science·Michael M C Lai
Jul 31, 2013·Veterinary Microbiology·Udeni B R BalasuriyaN James MacLachlan
Nov 26, 2002·Trends in Cell Biology·Jean DubuissonDarius Moradpour
May 12, 2009·Nature Reviews. Microbiology·Stanley Perlman, Jason Netland
Mar 20, 2004·Nature Reviews. Microbiology·Karla KirkegaardWilliam T Jackson
Jul 23, 2009·Acta Crystallographica. Section D, Biological Crystallography·Ioannis ManolaridisPaul A Tucker
Feb 25, 2011·Journal of Virology·Marne C HagemeijerCornelis A M de Haan
Aug 27, 2010·Journal of Virology·Monique H VerheijeCornelis A M de Haan
Sep 7, 2007·Journal of Virology·Matthew P Taylor, Karla Kirkegaard
Jul 30, 2010·Journal of Virology·Helen L StokesStuart G Siddell
Sep 15, 2007·Journal of Virology·Jennifer S SparksMark R Denison
Aug 25, 2006·Journal of Virology·Stanley G SawickiStuart G Siddell
Nov 17, 2006·Journal of Virology·Chandra TanguduThomas Gallagher
Dec 21, 2007·Journal of Virology·Paul Targett-AdamsJohn McLauchlan
Dec 17, 2009·Journal of Virology·Marne C HagemeijerCornelis A M de Haan
Feb 26, 2010·Journal of Virology·Mark A ClementzSusan C Baker

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