The vhs1 mutant form of herpes simplex virus virion host shutoff protein retains significant internal ribosome entry site-directed RNA cleavage activity.
The virion host shutoff (vhs) protein of herpes simplex virus (HSV) triggers global shutoff of host protein synthesis and accelerated turnover of host and viral mRNAs during HSV infection. As well, it induces endoribonucleolytic cleavage of RNA substrates when produced in a rabbit reticulocyte lysate (RRL) in vitro translation system. The vhs1 point mutation (Thr 214-->Ile) eliminates vhs function during virus infection and in transiently transfected mammalian cells and was therefore previously considered to abolish vhs activity. Here we demonstrate that the vhs1 mutant protein induces readily detectable endoribonuclease activity on RNA substrates bearing the internal ribosome entry site of encephalomyocarditis virus in the RRL assay system. These data document that the vhs1 mutation does not eliminate catalytic activity and raise the possibility that the vhs-dependent endoribonuclease employs more than one mode of substrate recognition.
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Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta, Canada T6G 2H7.
Saffran
H A
HA
Smiley
J R
JR
D000818:Animals; D004722:Endoribonucleases / Q000378:metabolism; D018259:Herpesvirus 1, Human / Q000378:metabolism* / Q000472:pathogenicity; D017354:Point Mutation; D012333:RNA, Messenger / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism; D011817:Rabbits; D012156:Reticulocytes; D012260:Ribonucleases; D012270:Ribosomes / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism*; D014771:Virion
D012333:RNA, Messenger; D012367:RNA, Viral; D014764:Viral Proteins; C070722:virion host shutoff protein, Simplexvirus; D004722:Endoribonucleases; D012260:Ribonucleases