Brefeldin A (BFA) was found to interfere with specific events of human cytomegalovirus (HCMV) maturation in human fibroblasts. Ultrastructural as well as biochemical studies suggested that short-term exposure of infected cultures to BFA during the late infectious cycle primarily prevented Golgi-dependent processes, e.g. envelopment of naked cytoplasmic nucleocapsids in the trans-Golgi network (TGN) and normal processing of glycoprotein B. In contrast, the nuclear phase of viral morphogenesis, e.g. transport budding at the nuclear envelope, was not impaired. These observations were compatible with the interpretation that HCMV morphogenesis may involve sequential budding events at the nuclear envelope and at cisternae of the TGN. BFA treatment during the early infectious cycle efficiently inhibited HCMV-DNA synthesis and thus late viral functions, preventing production of viral progeny. Cytotoxicity was excluded as a cause for these findings.
BonnerW. M.,
LaskeyR. A.1974; A film detection method for tritium-labeled proteins and nucleic acids in polyacrylamide gels. European Journal of Biochemistry 46:83–88
BrittW.,
VuglerL.,
ButfiloskiE.,
StephensE.1990; Cell surface expression of human cytomegalovirus (HCMV) gp 55–116 (gB): use of HCMV-recombinant vaccinia virus-infected cells in analysis of the human neutralizing antibody response. Journal of Virology 64:1079–1085
Campadelli-FiumeG.,
FarabegoliF.,
Di GaetaS.,
RoizmanB.1991; Origin of unenveloped capsids in the cytoplasm of cells infected with herpes simplex virus 1. Journal of Virology 65:1589–1595
ChangC. P.,
VesoleD. H.,
NelsonJ.,
OldstoneM. B. A.,
StinskiM.1989; Identification and expression of a human cytomegalovirus early glycoprotein. Journal of Virology 63:3330–3337
ChegeN. W.,
PfefferS. R.1990; Compartmentation of the Golgi complex: brefeldin A distinguishes trans-Golgi cisternae from the trans-Golgi network. Journal of Cell Biology 111:893–899
CheungP.,
BanfieldB. W.,
TufaroF.1991; Brefeldin A arrests the maturation and egress of herpes simplex virus particles during infection. Journal of Virology 65:1893–1904
CranageM.,
KouzaridesT.,
BankierA.,
SatchwellS.,
WestonK.,
TomlinsonP.,
BarrellB.,
HartH.,
BellS.,
MinsonA.,
SmithG.1986; Identification of the human cytomegalovirus glycoprotein B gene and induction of neutralizing antibodies via its expression in recombinant vaccina virus. EMBO Journal 5:3057–3063
DudenR.,
GriffithsG.,
FrankR.,
ArgosP.,
KreisTh.
1991; β-COP, a 110 kd protein associated with non-clathrin-coated vesicles and the Golgi complex, shows homology to β-adaptin. Cell 64:649–665
FujiwaraT.,
OdaK.,
YokotaS.,
TakatsukiA.,
IkeharaY.1988; Brefeldin A causes disassembly of the Golgi complex and accumulation of secretory proteins in the endoplasmic reticulum. Journal of Biological Chemistry 263:1845–1852
FurukawaT.,
FiorettiA.,
PlotkinS.1973; Growth characteristics of cytomegalovirus in human fibroblasts with the demonstration of protein synthesis early in viral replication. Journal of Virology 11:991–997
GretchD.,
KariB.,
RasmussenL.,
GehrzR.,
StinskiM.1988; Identification and characterization of three distinct families of glycoprotein complexes in the envelopes of human cytomegalovirus. Journal of Virology 62:875–881
HiraiK.,
WatanabeY.1976; Induction of alpha-type DNA polymerases in human cytomegalovirus-infected Wi-38 cells. Bio-chimica et biophysica acta 447:328–339
JonesF.,
GroseC.1988; Role of cytoplasmic vacuoles in varicella-zoster virus glycoprotein trafficking and virion envelopment. Journal of Virology 62:2701–2711
KariB.,
LussenhopN.,
GoertzR.,
Wabuke-BunotiM.,
RadekeR.,
GehrzR.1986; Characterization of monoclonal antibodies reactive to several biochemically distinct human cytomegalovirus glycoprotein complexes. Journal of Virology 60:345–352
KniessN.,
MachM.,
FayJ.,
BrittW.1991; Distribution of linear antigenic sites on glycoprotein gp 55 of human cytomegalovirus. Journal of Virology 65:138–146
LehnerR.,
MeyerH.,
MachM.1989; Identification and characterization of a human cytomegalovirus gene coding for a membrane protein that is conserved among human herpesviruses. Journal of Virology 63:3792–3800
Lippincott-SchwartzJ.,
DonaldsonJ. G.,
SchweizerA.,
BergerE. G.,
HauriH. P.,
YuanL. C.,
KlausnerR. D.1990; Microtubule-dependent retrograde transport of proteins into the ER in the presence of brefeldin A suggests an ER recycling pathway. Cell 60:821–836
LowryO. H.,
RosebroughN. J.,
FarrA. L.,
RandallR. J.1951; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
OrciL.,
TagayaM.,
AmherdtM.,
PerreletA.,
DonaldsonJ.,
Lippincott-SchwartzJ.,
KlausnerR.,
RothmanJ.1991; Brefeldin A, a drug that blocks secretion, prevents the assembly of non-clathrin-coated buds on Golgi cisternae. Cell 64:1183–1195
RadsakK.1978; Effect of herpes simplex virus type 1 infection on the cellular DNA polymerase activities of mouse cell cultures. Journal of General Virology 41:479–491
RadsakK. D.,
WederD.1981; Effect of 2-deoxy-D-glucose on cytomegalovirus-induced DNA synthesis in human fibroblasts. Journal of General Virology 57:33–42
RadsakK.,
BrücherK. H.,
BrittW.,
ShiouH.,
SchneiderD.,
KollertA.1990; Nuclear compartmentation of glycoprotein B of human cytomegalovirus. Virology 177:515–522
RasmussenL.,
NelsonM.,
NeffM.,
MeriganT.1988; Characterization of two different human cytomegalivirus glycoproteins which are targets for vims neutralizing antibody. Virology 163:308–318
RoizmanB.,
SearsA.1990; Herpes simplex viruses and their replication. In Virology 2nd edn pp. 1795–1841 Edited by
FieldsB. N.,
KnipeD. M.
New York: Raven Press;
SaitoM.,
SaitoM.,
RosenbergA.1984; Action of monensin, a monovalent cationophore, on cultured human fibroblasts: evidence that it induces high cellular accumulation of glucosyl- and lactosylceramide (gluco- and lactocerebroside). Biochemistry 23:1043–1046
SeveriB.,
LandiniM.,
GovoniE.1988; Human cytomegalovirus morphogenesis: an ultrastructural study of the late cytoplasmic phases. Archives of Virology 98:51–64
UlmerJ. B.,
PaladeG. E.1991; Effects of brefeldin A on the Golgi complex, endoplasmic reticulum and viral envelope glycoproteins in murine erythroleukemia cells. European Journal of Cell Biology 54:38–54
UtzU.,
BrittW.,
VuglerL.,
MachM.1989; Identification of a neutralizing epitope on glycoprotein gp58 of human cytomegalovirus. Journal of Virology 63:1995–2001
WhealyM. E.,
CardJ. P.,
MeadeR. P.,
RobbinsA. K.,
EnquistL. W.1991; Effect of brefeldin A on alphaherpesvirus membrane protein glycosylation and virus egress. Journal of Virology 65:1066–1081