Proteasome inhibitor MG132 blocks viral DNA replication and assembly of human cytomegalovirus.
This study provides evidence that proteasomal activity is required at multiple steps in human cytomegalovirus replication. Electron microscopy revealed that no viral particles were assembled in the presence of proteasome inhibitor MG132. Immunofluorescence and Western blot analyses using MG132 demonstrated that immediate early gene expression was suppressed at low but not high MOI. In contrast, expression of late proteins was completely blocked independent of MOI. Additionally, pulsed-field gel electrophoresis demonstrated that MG132 interferes with cleavage of HCMV DNA. Bromodeoxyuridine incorporation studies showed that de novo viral DNA synthesis is reduced in the presence of MG132. Furthermore, in contrast to previous hypotheses we demonstrated that neither the ND10 components PML and hDaxx nor NFkappaB activation represent the target for MG132.
Kaspari
Marion
M
Institute of Virology, Charité Campus Mitte, Universitätsmedizin Berlin, Charitéplatz 1, Berlin, Germany.
Tavalai
Nina
N
Stamminger
Thomas
T
Zimmermann
Albert
A
Schilf
Rita
R
Bogner
Elke
E
D000998:Antiviral Agents / Q000494:pharmacology; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000187:drug effects* / Q000502:physiology*; D003586:Cytomegalovirus Infections / Q000821:virology; D044708:DNA, Concatenated / Q000378:metabolism; D004279:DNA, Viral / Q000096:biosynthesis*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000821:virology; D006801:Humans; D007976:Leupeptins / Q000494:pharmacology*; D016328:NF-kappa B / Q000172:deficiency / Q000378:metabolism; D061988:Proteasome Inhibitors; D014764:Viral Proteins / Q000378:metabolism; D014771:Virion / Q000187:drug effects / Q000378:metabolism; D019065:Virus Assembly / Q000187:drug effects*; D014779:Virus Replication / Q000187:drug effects*
D000998:Antiviral Agents; D044708:DNA, Concatenated; D004279:DNA, Viral; D007976:Leupeptins; D016328:NF-kappa B; D061988:Proteasome Inhibitors; D014764:Viral Proteins; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde