Role of protein kinase A and the serine-rich region of herpes simplex virus type 1 ICP4 in viral replication

Efficient expression of herpes simplex virus genes requires the synthesis of functional ICP4, a nuclear phosphoprotein that contains a prominent serine-rich region between amino acids 142 and 210. Residues in this region not only are potential sites for phosphorylation but also are involved in the functions of ICP4. By comparing the growth of a virus in which this region is deleted (d8-10) with wild-type virus (KOS) in PC12 cells or PC12 cells that are deficient in cyclic AMP-dependent protein kinase (PKA), two observations were made: (i) the growth of wild-type virus was impaired by 1 to 2 orders of magnitude in the PKA-deficient cells, indicating the involvement of PKA in the growth cycle of herpes simplex virus type 1, and (ii) while the growth of d8-10 was impaired by almost 2 orders of magnitude in wild-type cells, it was not further impaired (as was that of wild-type virus) in PKA-deficient cells, implicating the region deleted in d8-10 as a possible target for cellular PKA. In trigeminal'ganglia of mice, the d8-10 mutant virus grew poorly; however, it established latency in nearly 90% of ganglia tested. Studies of the phosphorylation of wild-type and d8-10 ICP4 proteins revealed that the serine-rich region is a major determinant for phosphorylation of ICP4 in vivo and that the phosphorylation state could change as a function of the PKA activity. Consistent with this observation, the serine-rich region of ICP4 was shown to be a target for PKA in vitro. While intact ICP4 was readily phosphorylated by ICP4 in vitro, the d8-10 mutant ICP4 was not. Moreover, a synthethic peptide representing a sequence in the serine tract that is predicted to be a substrate for PKA was phosphorylated by PKA in vitro, having a Km within the physiological range. These data suggest that PKA plays a role in viral growth through phosphorylation of one or more sites on the ICP4 molecule.

[1]  N. DeLuca,et al.  Analysis of phosphorylation sites of herpes simplex virus type 1 ICP4 , 1996, Journal of virology.

[2]  N. DeLuca,et al.  Repression of activator-mediated transcription by herpes simplex virus ICP4 via a mechanism involving interactions with the basal transcription factors TATA-binding protein and TFIIB , 1995, Molecular and cellular biology.

[3]  M. Kretzschmar,et al.  A novel mediator of class II gene transcription with homology to viral immediate-early transcriptional regulators , 1994, Cell.

[4]  R. Roeder,et al.  Purification, cloning, and characterization of a human coactivator, PC4, that mediates transcriptional activation of class II genes , 1994, Cell.

[5]  C. Smith,et al.  ICP4, the major transcriptional regulatory protein of herpes simplex virus type 1, forms a tripartite complex with TATA-binding protein and TFIIB , 1993, Journal of virology.

[6]  D. Ginty,et al.  Nerve growth factor-induced neuronal differentiation after dominant repression of both type I and type II cAMP-dependent protein kinase activities. , 1991, The Journal of biological chemistry.

[7]  B. Roizman,et al.  ICP4, the major regulatory protein of herpes simplex virus, shares features common to GTP-binding proteins and is adenylated and guanylated , 1991, Journal of virology.

[8]  A. Papavassiliou,et al.  Analysis of the herpes simplex virus type 1 promoter controlling the expression of UL38, a true late gene involved in capsid assembly , 1991, Journal of virology.

[9]  S. Silverstein,et al.  The interaction of ICP4 with cell/infected‐cell factors and its state of phosphorylation modulate differential recognition of leader sequences in herpes simplex virus DNA. , 1991, The EMBO journal.

[10]  N. DeLuca,et al.  A second-site revertant of a defective herpes simplex virus ICP4 protein with restored regulatory activities and impaired DNA-binding properties , 1991, Journal of virology.

[11]  N. DeLuca,et al.  Activities of heterodimers composed of DNA-binding- and transactivation-deficient subunits of the herpes simplex virus regulatory protein ICP4 , 1991, Journal of virology.

[12]  R. Everett,et al.  A prominent serine-rich region in Vmw175, the major transcriptional regulator protein of herpes simplex virus type 1, is not essential for virus growth in tissue culture. , 1990, The Journal of general virology.

[13]  N. DeLuca,et al.  Functional relevance of specific interactions between herpes simplex virus type 1 ICP4 and sequences from the promoter-regulatory domain of the viral thymidine kinase gene , 1990, Journal of virology.

[14]  D. Baltimore,et al.  Activation in vitro of NF-κB" by phosphorylation of its inhibitor IκB" , 1990, Nature.

[15]  E. Krebs,et al.  Myb DNA binding inhibited by phosphorylation at a site deleted during oncogenic activation , 1990, Nature.

[16]  B. Roizman,et al.  Binding of the herpes simplex virus major regulatory protein to viral DNA. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[17]  N. DeLuca,et al.  Separation of primary structural components conferring autoregulation, transactivation, and DNA-binding properties to the herpes simplex virus transcriptional regulatory protein ICP4 , 1989, Journal of virology.

[18]  D. O’Callaghan,et al.  DNA sequence and comparative analyses of the equine herpesvirus type 1 immediate early gene. , 1989, Virology.

[19]  A. Cheung DNA nucleotide sequence analysis of the immediate-early gene of pseudorabies virus. , 1989, Nucleic acids research.

[20]  C. Denis,et al.  Cyclic AMP-dependent protein kinase phosphorylates and inactivates the yeast transcriptional activator ADR1 , 1989, Cell.

[21]  P. Kattar-Cooley,et al.  Characterization of the DNA-binding properties of herpes simplex virus regulatory protein ICP4 , 1989, Journal of virology.

[22]  K. Tyler,et al.  Immediate-early regulatory gene mutants define different stages in the establishment and reactivation of herpes simplex virus latency , 1989, Journal of virology.

[23]  R. Everett,et al.  The regions of the herpes simplex virus type 1 immediate early protein Vmw175 required for site specific DNA binding closely correspond to those involved in transcriptional regulation. , 1988, Nucleic acids research.

[24]  D. Walsh,et al.  The catalytic subunit of cAMP-dependent protein kinase induces expression of genes containing cAMP-responsive enhancer elements , 1988, Nature.

[25]  M. Ptashne How eukaryotic transcriptional activators work , 1988, Nature.

[26]  R. Everett,et al.  Mutational dissection of the HSV-1 immediate-early protein Vmw175 involved in transcriptional transactivation and repression. , 1988, Virology.

[27]  B. Roizman,et al.  The DNA-binding properties of the major regulatory protein alpha 4 of herpes simplex viruses. , 1988, Science.

[28]  N. DeLuca,et al.  Physical and functional domains of the herpes simplex virus transcriptional regulatory protein ICP4 , 1988, Journal of virology.

[29]  N. DeLuca,et al.  Activities of herpes simplex virus type 1 (HSV-1) ICP4 genes specifying nonsense peptides. , 1987, Nucleic acids research.

[30]  M. Muller Binding of the herpes simplex virus immediate-early gene product ICP4 to its own transcription start site , 1987, Journal of virology.

[31]  K. Struhl,et al.  Functional dissection of a eukaryotic transcriptional activator protein, GCN4 of Yeast , 1986, Cell.

[32]  K. Wilcox,et al.  Association of the herpes simplex virus regulatory protein ICP4 with specific nucleotide sequences in DNA. , 1986, Nucleic acids research.

[33]  R. Tjian,et al.  Affinity purification of sequence-specific DNA binding proteins. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[34]  B. Roizman,et al.  DNA-binding site of major regulatory protein alpha 4 specifically associated with promoter-regulatory domains of alpha genes of herpes simplex virus type 1. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[35]  B. Roizman,et al.  Alpha 4, the major regulatory protein of herpes simplex virus type 1, is stably and specifically associated with promoter-regulatory domains of alpha genes and of selected other viral genes. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[36]  D. McGeoch,et al.  Complete DNA sequence of the short repeat region in the genome of herpes simplex virus type 1. , 1986, Nucleic acids research.

[37]  N. DeLuca,et al.  Isolation and characterization of deletion mutants of herpes simplex virus type 1 in the gene encoding immediate-early regulatory protein ICP4 , 1985, Journal of virology.

[38]  N. DeLuca,et al.  Activation of immediate-early, early, and late promoters by temperature-sensitive and wild-type forms of herpes simplex virus type 1 protein ICP4 , 1985, Molecular and cellular biology.

[39]  H. Cheng,et al.  Circular dichroic evidence for an ordered sequence of ligand/binding site interactions in the catalytic reaction of the cAMP-dependent protein kinase. , 1985, Biochemistry.

[40]  N. DeLuca,et al.  Temperature-sensitive mutants in herpes simplex virus type 1 ICP4 permissive for early gene expression , 1984, Journal of virology.

[41]  B. Roizman,et al.  Characterization of herpes simplex virus 1 alpha proteins 0, 4, and 27 with monoclonal antibodies , 1984, Journal of virology.

[42]  C. Preston,et al.  Poly(ADP-ribosyl)ation of a herpes simplex virus immediate early polypeptide. , 1983, Virology.

[43]  R. Dixon,et al.  Fine-structure mapping and functional analysis of temperature-sensitive mutants in the gene encoding the herpes simplex virus type 1 immediate early protein VP175 , 1980, Journal of virology.

[44]  R. Watson,et al.  A herpes simplex virus type 1 function continuously required for early and late virus RNA synthesis , 1980, Nature.

[45]  B. Roizman,et al.  Herpes simplex virus phosphoproteins. I. Phosphate cycles on and off some viral polypeptides and can alter their affinity for DNA , 1980, Journal of virology.

[46]  C. Preston Abnormal properties of an immediate early polypeptide in cells infected with the herpes simplex virus type 1 mutant tsK , 1979, Journal of virology.

[47]  C. Preston Control of herpes simplex virus type 1 mRNA synthesis in cells infected with wild-type virus or the temperature-sensitive mutant tsK , 1979, Journal of virology.

[48]  A. Davison,et al.  Recombinants between herpes simplex virus types 1 and 2: analyses of genome structures and expression of immediate early polypeptides , 1978, Journal of virology.

[49]  B. Roizman,et al.  Molecular genetics of herpes simplex virus: demonstration of regions of obligatory and nonobligatory identity within diploid regions of the genome by sequence replacement and insertion. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[50]  E. Krebs,et al.  Role of multiple basic residues in determining the substrate specificity of cyclic AMP-dependent protein kinase. , 1977, The Journal of biological chemistry.

[51]  M. Wolff,et al.  Regulation of herpesvirus macromolecular synthesis. V. Properties of alpha polypeptides made in HSV-1 and HSV-2 infected cells. , 1977, Virology.

[52]  B. Roizman,et al.  Regulation of herpesvirus macromolecular synthesis: sequential transition of polypeptide synthesis requires functional viral polypeptides. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[53]  R. Courtney,et al.  Isolation and characterization of a large molecular-weight polypeptide of herpes simplex virus type 1. , 1974, Virology.

[54]  B. Roizman,et al.  Regulation of Herpesvirus Macromolecular Synthesis I. Cascade Regulation of the Synthesis of Three Groups of Viral Proteins , 1974, Journal of virology.

[55]  B. Roizman,et al.  Proteins Specified by Herpes Simplex Virus VIII. Characterization and Composition of Multiple Capsid Forms of Subtypes 1 and 2 , 1972, Journal of virology.

[56]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[57]  P. Spear,et al.  Proteins spcified by herpes simplex virus. II. Viral glycoprotins associated with cellular membranes. , 1970, Journal of virology.

[58]  R. Pearson,et al.  Protein kinase phosphorylation site sequences and consensus specificity motifs: tabulations. , 1991, Methods in enzymology.

[59]  A. Edelman,et al.  Protein serine/threonine kinases. , 1987, Annual review of biochemistry.

[60]  R. Roskoski [1] Assays of protein kinase , 1983 .

[61]  R. Roskoski Assays of protein kinase. , 1983, Methods in enzymology.

[62]  P. Y. Chou,et al.  Empirical predictions of protein conformation. , 1978, Annual review of biochemistry.