The virion-packaged endoribonuclease of herpes simplex virus 1 cleaves mRNA in polyribosomes.
The virion host shutoff protein product of the U(L)41 gene of herpes simplex virus 1 is an endoribonuclease that selectively degrades mRNAs during the first hours after infection. Specifically, in contrast to the events in uninfected cells or cells infected with a mutant lacking the RNase, in wild-type virus-infected cells mRNA of housekeeping genes exemplified by GAPDH is degraded rapidly, whereas mRNAs containing AU elements are cleaved and the 5' cleavage product of these RNAs persists for many hours. We report that in wild-type virus-infected cells there was a rapid increase in the number and size of processing bodies (P-bodies). These P-bodies were also preset in cycloheximide (CHX)-treated cells but not in either treated or untreated uninfected cells or cells infected with the RNase minus mutant. Additional studies revealed that polyribosomes extracted from cytoplasm of wild-type virus-infected cells treated with CHX and displayed in sucrose gradients contained ribosome-loaded, truncated AU-rich mRNAs lacking the 3' UTR and poly(A) tails. The results suggest that the virion RNase is bound to polyribosomes by virtue of the reported association with translation machinery and cleaves the RNAs 5' to the AU elements. In contrast to the slow degradation of the of the residual 5' domain, the 3' UTR of the AU-rich mRNA and the GAPDH mRNA are rapidly degraded in wild-type virus-infected cells.
Taddeo
Brunella
B
The Marjorie B. Kovler Viral Oncology Laboratories, University of Chicago, Chicago, IL 60637, USA.
Zhang
Weiran
W
Roizman
Bernard
B
D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D022083:Cytoplasmic Structures / Q000187:drug effects / Q000473:pathology / Q000821:virology; D004722:Endoribonucleases / Q000378:metabolism*; D006367:HeLa Cells; D006561:Herpes Simplex / Q000821:virology; D018259:Herpesvirus 1, Human / Q000201:enzymology* / Q000502:physiology*; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D008954:Models, Biological; D011132:Polyribosomes / Q000187:drug effects / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012260:Ribonucleases; D013997:Time Factors; D014764:Viral Proteins / Q000378:metabolism*; D014771:Virion / Q000187:drug effects / Q000201:enzymology*; D019065:Virus Assembly / Q000187:drug effects
D051017:Apoptosis Regulatory Proteins; C099815:IER3 protein, human; D008565:Membrane Proteins; D012333:RNA, Messenger; D014764:Viral Proteins; C070722:virion host shutoff protein, Simplexvirus; D003513:Cycloheximide; D004722:Endoribonucleases; D012260:Ribonucleases