title: Change of processing and nucleocytoplasmic transport of mRNA in HSV-1-infected cells.
abstract: A monoclonal antibody (MAb) raised against the pore-complex lamina fraction from CV-1 cells was used to study alterations of gene expression in herpes simplex virus type 1 (HSV-1)-infected cells. This MAb, which recognized only one cellular polypeptide of 60,000 Da, selectively stained the nucleus in immunofluorescence microscopy, showing a punctuated pattern either at the nuclear surface or at the nuclear rim. By immunoelectron microscopy, the p60 antigen could be localized in the nuclear pore complex structure. Infection of CV-1 cells with HSV-1 resulted in a drastic change of the nuclear staining pattern. Four hours p.i., a clustering of the p60 antigen and, 12 h p.i., a formation of finger-like holes, penetrating the nucleus, occurred. Later in infection (22 h p.i.) the antigen was found to be almost absent. By RNA blot hybridization it was demonstrated that, after HSV-1 infection, the level of cellular mRNA (beta-tubulin) gradually decreased, while the level of HSV major DNA binding protein (DBP) mRNA increased, reaching maximal level 3-6 h p.i. Interestingly, the level of beta-tubulin gene transcripts changed differentially in the polysomal and in the nuclear fraction during the initial phase of infection, in contrast to the viral DBP transcripts, indicating that, after HSV infection, host cell transcripts accumulate in the nucleus. Evidence is presented indicating that this change is not due to altered nucleocytoplasmic mRNA transport but is due to an impaired splicing of host cell transcripts in HSV-infected cells. The MAb, directed against the nuclear pore p60 antigen, strongly inhibited the ATP-dependent efflux of both cellular and viral mRNA from isolated nuclei. The ATP-dependence of the efflux did not change during viral infection. However, the inhibitory potency of the MAb was found to be lost at the final stage of HSV infection, paralleling the loss of p60 antigen.
authors.affiliation: Institut für Physiologische Chemie, Abteilung Angewandte Molekularbiologie, Mainz, F.R.G.
authors.forename: H C
authors.initials: HC
authors.lastname: Schröder
authors.forename: D
authors.initials: D
authors.lastname: Falke
authors.forename: K
authors.initials: K
authors.lastname: Weise
authors.forename: M
authors.initials: M
authors.lastname: Bachmann
authors.forename: M
authors.initials: M
authors.lastname: Carmo-Fonseca
authors.forename: T
authors.initials: T
authors.lastname: Zaubitzer
authors.forename: W E
authors.initials: WE
authors.lastname: Müller
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D000911:Antibodies, Monoclonal; D000956:Antigens, Viral / Q000235:genetics; D001693:Biological Transport, Active / Q000187:drug effects; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000276:immunology; D005786:Gene Expression Regulation; D012323:RNA Processing, Post-Transcriptional; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism*; D018139:Simplexvirus / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014158:Transcription, Genetic; D014404:Tubulin / Q000235:genetics / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; D014404:Tubulin; D000255:Adenosine Triphosphate