Herpes simplex virus inhibits host cell splicing, and regulatory protein ICP27 is required for this effect.
While the majority of metazoan genes and those of the DNA viruses which infect them contain introns which require RNA splicing, herpes simplex virus type 1 (HSV-1) encodes relatively few spliced products. We previously showed that the HSV-1 immediate-early protein ICP27 decreased the levels of spliced target mRNAs in transfections and spliced cellular mRNAs during infection, suggesting that ICP27 may function in impairing host cell splicing. Here, we show that during infections with the wild type, but not in infections with an ICP27 viral mutant termed 27-LacZ, precursor RNA accumulated for a virus transcript which contained introns. Pre-mRNA accumulation in the nucleus was greater than that in the cytoplasm, indicating that splicing rather than transport was affected. Furthermore, splicing of a beta-globin pre-mRNA substrate was inhibited in nuclear extracts from wild-type-infected cells but not in extracts from cells infected with 27-LacZ. The inhibitory activity in extracts from wild-type-infected cells was able to reduce the splicing efficiency of competent extracts in biochemical complementation assays. ICP27 appeared to be responsible for this decrease, because the splicing activity of an extract from cells infected with an ICP27 ts mutant was significantly reduced after incubation of the extract at the permissive temperature to allow renaturation of the conformationally defective ICP27 protein. These results strongly suggest that HSV-1 infection inhibits host cell splicing through the action of ICP27.
Hardy
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WR
Department of Microbiology and Molecular Genetics, College of Medicine, University of California, Irvine 92717-4025.
Sandri-Goldin
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RM
D002478:Cells, Cultured; D003001:Cloning, Molecular; D005347:Fibroblasts; D005914:Globins / Q000096:biosynthesis*; D006367:HeLa Cells; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000502:physiology*; D006801:Humans; D017874:Immediate-Early Proteins; D010957:Plasmids; D012322:RNA Precursors / Q000378:metabolism*; D012326:RNA Splicing; D012367:RNA, Viral / Q000096:biosynthesis; D012097:Repressor Proteins / Q000378:metabolism*; D012867:Skin; D014162:Transfection; D014764:Viral Proteins / Q000096:biosynthesis / Q000378:metabolism*
C057120:ICP27 protein, human herpesvirus 1; D017874:Immediate-Early Proteins; D012322:RNA Precursors; D012367:RNA, Viral; D012097:Repressor Proteins; D014764:Viral Proteins; D005914:Globins