U(S)3 and U(S)3.5 protein kinases of herpes simplex virus 1 differ with respect to their functions in blocking apoptosis and in virion maturation and egress.
Previously, we reported that the U(S)3 protein kinase blocks apoptosis, that it activates protein kinase A (PKA), that activation of PKA blocks apoptosis in cells infected with a U(S)3 deletion mutant, and that an overlapping transcriptional unit encodes a truncated kinase designated U(S)3.5. Here, we report the properties of the kinases based on comparisons of herpes simplex virus and baculoviruses expressing U(S)3 or U(S)3.5 kinase. Specifically, we report the following. (i) Both kinases mediate the phosphorylation of HDAC1, HDAC2, and the PKA regulatory IIalpha subunit in the absence of other viral proteins. (ii) Both enzymes mediate the phosphorylation of largely identical sets of proteins carrying the phosphorylation consensus site of PKA, but only U(S)3 blocks apoptosis, suggesting that it is U(S)3 and not PKA that is responsible for the phosphorylation of the proteins bearing the shared consensus phosphorylation site and the antiapoptotic activity. (iii) Both kinases cofractionate with mitochondria. Immune depletion of the U(S)3 and U(S)3.5 kinases from the cytoplasm removed the kinases from the supernatant fraction, but not from the mitochondrial fraction, and therefore, if the antiapoptotic activity of the U(S)3 kinase is expressed in mitochondria, the localization signal and the antiapoptotic functions are located on different parts of the protein. (iv) The U(S)3 protein kinase is required for the translocation of virus particles from the nucleus. Although the U(L)31 protein is phosphorylated in cells infected with the mutant expressing U(S)3.5 kinase, the release of virus particles from nuclei was impeded in some cells, suggesting that the U(S)3 kinase affects the modification of the nuclear membrane more efficiently than the U(S)3.5 kinase.
Poon
Alice P W
AP
The Marjorie B. Kovler Viral Oncology Laboratories, The University of Chicago, 910 East 58th Street, Chicago, Illinois 60637, USA.
Benetti
Luca
L
Roizman
Bernard
B
D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D002467:Cell Nucleus / Q000821:virology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D018259:Herpesvirus 1, Human / Q000502:physiology*; D056464:Histone Deacetylase 2; D006655:Histone Deacetylases / Q000378:metabolism; D006801:Humans; D017874:Immediate-Early Proteins / Q000502:physiology; D007976:Leupeptins / Q000494:pharmacology; D009687:Nuclear Proteins / Q000378:metabolism; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D017346:Protein Serine-Threonine Kinases / Q000502:physiology*; D011817:Rabbits; D012097:Repressor Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism / Q000502:physiology*; D054334:Viral Regulatory and Accessory Proteins; D014771:Virion / Q000502:physiology*; D051025:bcl-Associated Death Protein / Q000502:physiology
C057294:ICP22 protein, human herpesvirus 1; D017874:Immediate-Early Proteins; D007976:Leupeptins; D009687:Nuclear Proteins; D012097:Repressor Proteins; C108721:UL31 protein, Human herpesvirus 1; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; D051025:bcl-Associated Death Protein; D017346:Protein Serine-Threonine Kinases; C095207:US3 protein, Human herpesvirus 1; D017868:Cyclic AMP-Dependent Protein Kinases; D056464:Histone Deacetylase 2; D006655:Histone Deacetylases; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde