Analysis of herpes simplex virus-induced mRNA destabilizing activity using an in vitro mRNA decay system.
Most host mRNAs are degraded soon after infection of cells with herpes simplex virus type 1 (HSV-1). This early shutoff or early destabilization response is induced by a virion component, the virion host shutoff (vhs) protein. HSV-1 mutants, vhs1 and vhs-delta Sma, which produce defective or inactive vhs protein, fail to induce early shutoff. We have used an in vitro mRNA decay system to analyze the destabilization process. Polysomes from uninfected human erythroleukemia cells, used as a source of target mRNAs, were mixed with polysomes or with post-polysomal supernatant (S130) from HSV-1- or mock-infected murine erythroleukemia cells. Normally stable gamma-globin mRNA was destabilized by approximately 15-fold with S130 from wild-type virus-infected cells but was not destabilized with S130 from mock-infected cells or from cells infected with either of the two HSV mutants. The virus-induced destabilizing activity had no significant effect on the in vitro half-lives of two normally unstable mRNAs, histone and c-myc. No destabilizing activity was detected in polysomes from infected cells. We conclude that a virus-induced destabilizer activity can function in vitro, is located in the S130 of infected cells, and accelerates the decay rates of some, but not all, polysome-associated host mRNAs.
Sorenson
C M
CM
McArdle Laboratory for Cancer Research, Madison, WI 53706.
Hart
P A
PA
Ross
J
J
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D002460:Cell Line; D005914:Globins / Q000235:genetics; D006801:Humans; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D011132:Polyribosomes / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D012260:Ribonucleases; D016415:Sequence Alignment; D018139:Simplexvirus / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000378:metabolism*
D012333:RNA, Messenger; D014764:Viral Proteins; C070722:virion host shutoff protein, Simplexvirus; D005914:Globins; D012260:Ribonucleases