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Journal of Virology, November 1999, p. 9555-9567, Vol. 73, No. 11
Sir Albert Sakzewski Virus Research Centre,
Royal Children's Hospital, Herston, Brisbane, Australia
4029,1 and Centre for Microscopy and
Microanalysis, University of Queensland, St. Lucia, Brisbane, Australia
40722
Received 19 April 1999/Accepted 23 July 1999
Replication of the flavivirus Kunjin virus is associated with
virus-induced membrane structures within the cytoplasm of infected cells; these membranes appear as packets of vesicles associated with
the sites of viral RNA synthesis and as convoluted membranes (CM) and
paracrystalline arrays (PC) containing the components of the
virus-specified protease (E. G. Westaway, J. M. Mackenzie, M. T. Kenney, M. K. Jones, and A. A. Khromykh, J. Virol. 71:6650-6661, 1997). To determine the cellular origins of these
membrane structures, we compared the immunolabelling patterns of
several cell markers in relation to these sites by immunofluorescence
and immunoelectron microscopy. A marker for the trans-Golgi
membranes and the trans-Golgi network,
1,4-galactosyltransferase (GalT), was redistributed to large foci in
the cytoplasm of Kunjin virus-infected cells, partially coincident with
immunofluorescent foci associated with the putative sites of viral RNA
synthesis. As determined by immunoelectron microscopy, the induced
vesicle packets contained GalT, whereas the CM and PC contained a
specific protein marker for the intermediate compartment (ERGIC53). A
further indicator of the role of cellular organelles in their
biogenesis was the observation that the Golgi apparatus-disrupting
agent brefeldin A prevented further development of immunofluorescent
foci of induced membranes if added before the end of the latent period
but that once formed, these membrane foci were resistant to brefeldin A
dispersion. Reticulum membranes emanating from the induced CM and PC
were also labelled with the rough endoplasmic reticulum marker
anti-protein disulfide isomerase and were obviously redistributed
during infection. This is the first report identifying
trans-Golgi membranes and the intermediate compartment as
the apparent sources of the flavivirus-induced membranes involved in
events of replication.
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Markers for trans-Golgi Membranes and
the Intermediate Compartment Localize to Induced Membranes with
Distinct Replication Functions in Flavivirus-Infected Cells
*
Corresponding author. Mailing address: Sir Albert
Sakzewski Virus Research Centre, Royal Children's Hospital, Herston,
Brisbane, Australia 4029. Phone: 61 7 3253 1569. Fax: 61 7 3253 1401. E-mail: mackenzi{at}biosci.uq.oz.au.
SASVRC publication 100.
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