Gene expression and cytopathic effect of vaccinia virus inactivated by psoralen and long-wave UV light.
Induction of the cytopathic effect (CPE) in cells infected with poxvirus seems ubiquitous in that it has been associated with all different strains and preparations of poxviruses, regardless of the replicating status of these viruses. The study of the mechanisms by which CPE is induced by nonreplicating poxviruses is hampered by the lack of any noncytopathic mutant strains and preparations. In this paper, we report on the patterns of gene expression and induction of CPE by vaccinia viruses treated by limited cross-linking with psoralen and long-wave UV light (PLWUV). We show that treatment of cell-free virus with PLWUV could inactivate viral replication without abolishing the ability of the virus to infect cells. Viral transcription as indicated by reporter genes was generally enhanced and prolonged under early viral promoters and abolished under late promoters. Furthermore, increasing the levels of cross-linking with PLWUV resulted in a decrease and abolishment of viral expression of a large reporter gene and a concomitant loss of the induction of CPE. Cells infected with such a virus were able to express the reporter genes and proliferate. The generation of nonreplicating and noncytopathic recombinant vaccinia viruses may help in studies of the mechanisms of CPE induction by poxvirus and may facilitate the use of poxviral vectors in broader areas of research and clinical applications.
Department of Surgery, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
K
K
Tsung
J H
JH
Yim
W
W
Marti
R M
RM
Buller
J A
JA
Norton
D000818:Animals; D002460:Cell Line; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D003588:Cytopathogenic Effect, Viral / Q000187:drug effects* / Q000528:radiation effects; D004279:DNA, Viral / Q000378:metabolism; D005363:Ficusin / Q000494:pharmacology*; D015967:Gene Expression Regulation, Viral / Q000187:drug effects*; D017930:Genes, Reporter; D017319:Photosensitizing Agents / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D014466:Ultraviolet Rays; D014616:Vaccinia virus / Q000187:drug effects* / Q000235:genetics / Q000472:pathogenicity / Q000528:radiation effects; D014779:Virus Replication / Q000187:drug effects / Q000528:radiation effects
D003432:Cross-Linking Reagents; D004279:DNA, Viral; D017319:Photosensitizing Agents; D005363:Ficusin