title: Theiler's murine encephalomyelitis virus infection induces a redistribution of heat shock proteins 70 and 90 in BHK-21 cells, and is inhibited by novobiocin and geldanamycin.
abstract: Theiler's murine encephalomyelitis virus (TMEV) is a positive-sense RNA virus belonging to the Cardiovirus genus in the family Picornaviridae. In addition to other host cellular factors and pathways, picornaviruses utilise heat shock proteins (Hsps) to facilitate their propagation in cells. This study investigated the localisation of Hsps 70 and 90 in TMEV-infected BHK-21 cells by indirect immunofluorescence and confocal microscopy. The effect of Hsp90 inhibitors novobiocin (Nov) and geldanamycin (GA) on the development of cytopathic effect (CPE) induced by infection was also examined. Hsp90 staining was uniformly distributed in the cytoplasm of uninfected cells but was found concentrated in the perinuclear region during late infection where it overlapped with the signal for non-structural protein 2C within the viral replication complex. Hsp70 redistributed into the vicinity of the viral replication complex during late infection, but its distribution did not overlap with that of 2C. Inhibition of Hsp90 by GA and Nov had a negative effect on virus growth over a 48-h period as indicated by no observable CPE in treated compared to untreated cells. 2C was detected by Western analysis of GA-treated infected cell lysates at doses between 0.01 and 0.125 μM, suggesting that processing of viral precursors was not affected in the presence of this drug. In contrast, 2C was absent in cell lysates of Nov-treated cells at doses above 10 μM, although CPE was evident 48 hpi. This is the first study describing the dynamic behaviour of Hsps 70 and 90 in TMEV-infected cells and to identify Hsp90 as an important host factor in the life cycle of this virus.
authors: Department of Biochemistry, Microbiology and Biotechnology, Rhodes University, 6140, Grahamstown, South Africa.
authors: Lorraine Z
authors: LZ
authors: Mutsvunguma
authors: Boitumelo
authors: B
authors: Moetlhoa
authors: Adrienne L
authors: AL
authors: Edkins
authors: Garry A
authors: GA
authors: Luke
authors: Gregory L
authors: GL
authors: Blatch
authors: Caroline
authors: C
authors: Knox
mesh_terms: D000818:Animals; D016227:Benzoquinones / Q000378:metabolism* / Q000494:pharmacology; D018188:Cardiovirus Infections / Q000378:metabolism*; D002460:Cell Line / Q000187:drug effects / Q000821:virology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000378:metabolism* / Q000494:pharmacology; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D047029:Lactams, Macrocyclic / Q000378:metabolism* / Q000494:pharmacology; D009675:Novobiocin / Q000378:metabolism* / Q000494:pharmacology; D017945:Theilovirus / Q000187:drug effects / Q000472:pathogenicity / Q000502:physiology*; D014779:Virus Replication / Q000187:drug effects
chemical_list: D016227:Benzoquinones; D004791:Enzyme Inhibitors; D018840:HSP70 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins; D047029:Lactams, Macrocyclic; D009675:Novobiocin; C001277:geldanamycin