U(S)3 protein kinase of herpes simplex virus 1 blocks caspase 3 activation induced by the products of U(S)1.5 and U(L)13 genes and modulates expression of transduced U(S)1.5 open reading frame in a cell type-specific manner

J Virol. 2002 Jan;76(2):743-54. doi: 10.1128/jvi.76.2.743-754.2002.

Abstract

The coding domain of the herpes simplex virus type 1 (HSV-1) alpha22 gene encodes two proteins, the 420-amino-acid infected-cell protein 22 (ICP22) and U(S)1.5, a protein colinear with the carboxyl-terminal domain of ICP22. In HSV-1-infected cells, ICP22 and U(S)1.5 are extensively modified by the U(L)13 and U(S)3 viral protein kinases. In this report, we show that in contrast to other viral proteins defined by their properties as alpha proteins, U(S)1.5 becomes detectable and accumulated only at late times after infection. Moreover, significantly more U(S)1.5 protein accumulated in cells infected with a mutant lacking the U(L)13 gene than in cells infected with wild-type virus. To define the role of viral protein kinases on the accumulation of U(S)1.5 protein, rabbit skin cells or Vero cells were exposed to recombinant baculoviruses that expressed U(S)1.5, U(L)13, or U(S)3 proteins under a human cytomegalovirus immediate-early promoter. The results were as follows. (i) Accumulation of the U(S)1.5 protein was reduced by concurrent expression of the U(L)13 protein kinase and augmented by concurrent expression of the U(S)3 protein kinase. The magnitude of the reduction or increase in the accumulation of the U(S)1.5 protein was cell type dependent. The effect of U(L)13 kinase appears to be specific inasmuch as it did not affect the accumulation of glycoprotein D in cells doubly infected by recombinant baculoviruses expressing these genes. (ii) The reduction in accumulation of the U(S)1.5 protein was partially due to proteasome-dependent degradation. (iii) Both U(S)1.5 and U(L)13 proteins activated caspase 3, indicative of programmed cell death. (iv) Concurrent expression of the U(S)3 protein kinase blocked activation of caspase 3. The results are concordant with those published elsewhere (J. Munger and B. Roizman, Proc. Natl. Acad. Sci. USA 98:10410-10415, 2001) that the U(S)3 protein kinase can block apoptosis by degradation or posttranslational modification of BAD.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Baculoviridae / genetics
  • Caspase 3
  • Caspase Inhibitors*
  • Caspases / metabolism
  • Cell Line
  • Chlorocebus aethiops
  • Cysteine Endopeptidases / metabolism
  • Down-Regulation
  • Enzyme Activation
  • Gene Expression Regulation, Viral
  • Genes, Viral / genetics
  • Herpesvirus 1, Human / enzymology*
  • Herpesvirus 1, Human / genetics*
  • Immediate-Early Proteins / biosynthesis
  • Immediate-Early Proteins / chemistry
  • Immediate-Early Proteins / genetics
  • Immediate-Early Proteins / metabolism*
  • Kinetics
  • Models, Biological
  • Multienzyme Complexes / metabolism
  • Open Reading Frames / genetics*
  • Organ Specificity
  • Proteasome Endopeptidase Complex
  • Protein Kinases / genetics
  • Protein Kinases / metabolism*
  • Protein Processing, Post-Translational
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism*
  • Rabbits
  • Recombinant Proteins / metabolism
  • Vero Cells
  • Viral Envelope Proteins / metabolism
  • Viral Proteins*
  • Viral Regulatory and Accessory Proteins

Substances

  • Caspase Inhibitors
  • ICP22 protein, human herpesvirus 1
  • Immediate-Early Proteins
  • Multienzyme Complexes
  • Recombinant Proteins
  • Viral Envelope Proteins
  • Viral Proteins
  • Viral Regulatory and Accessory Proteins
  • glycoprotein D, Equid herpesvirus 1
  • EUS1 protein, Equine herpesvirus 1
  • Protein Kinases
  • UL13 protein, Simplexvirus
  • Protein Serine-Threonine Kinases
  • US3 protein, Human herpesvirus 1
  • CASP3 protein, human
  • Caspase 3
  • Caspases
  • Cysteine Endopeptidases
  • Proteasome Endopeptidase Complex