HERPES-SIMPLEX VIRUS TYPE-1 ICP0 REGULATES EXPRESSION OF IMMEDIATE-EARLY, EARLY, AND LATE GENES IN PRODUCTIVELY INFECTED-CELLS

被引:226
作者
CAI, WZ
SCHAFFER, PA
机构
[1] HARVARD UNIV,SCH MED,DANA FARBER CANC INST,TUMOR VIRUS GENET LAB,BOSTON,MA 02115
[2] HARVARD UNIV,SCH MED,DEPT MICROBIOL & MOLEC GENET,BOSTON,MA 02115
关键词
D O I
10.1128/JVI.66.5.2904-2915.1992
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The herpes simplex virus type 1 protein, ICP0, can activate expression of all kinetic classes of viral promoters in transient expression assays. To examine the role of ICP0 in the regulation of viral gene expression during productive infection, we characterized the wild-type virus, an ICP0 null mutant (7134), and several ICP0 nonsense mutant viruses with regard to virus replication and protein synthesis in Vero cells. Relative to wild-type virus, 7134 was severely deficient in viral growth and protein synthesis at low multiplicities of infection but exhibited a nearly wild-type phenotype at high multiplicities. The phenotypes of the ICP0 nonsense mutants were intermediate between those of the wild-type virus and 7134 in that the more ICP0-coding sequence expressed by a given nonsense mutant, the more wild type-like was its phenotype. The location of the ICP0 domain responsible for transactivation during productive infection was confirmed to be within the N-terminal portion of the protein, as previously shown in transient expression assays. Immunoprecipitation and immunofluorescence tests were used to detect low-level expression of selected immediate-early (IE), early (E), and late (L) proteins by mutant and wild-type viruses following low-multiplicity infection. The 7134 deletion mutant and several nonsense mutants expressed markedly reduced levels of E and L proteins but wild-type levels of the IE protein, ICP4. Because the latency-associated transcripts (LATs) are specified by the strand opposite that which encodes ICP0, the ICP0 deletion and nonsense mutants are by definition ICP0-LAT double mutants. The ability of a LAT-ICP0+ mutant to replicate as efficiently as wild-type virus at low multiplicities and the ability of ICP0-expressing 0-28 cells to complement the defects of the mutants in E and L protein synthesis indicates that the phenotypes of the mutants are caused by mutations in ICP0 and not the LATs. Thus, we conclude that ICP0 up-regulates E and L but not necessarily IE gene expression during productive infection. The activation of IE gene expression by ICP0 during productive infection is likely overshadowed by the activity of the virion-associated protein, VP16. This hypothesis was tested by transfection of Vero cells with infectious mutant and wild-type viral DNAs. In such tests, no VP16 is present at early times posttransfection. Significantly fewer cells transfected with infectious 7134 DNA expressed ICP4 than cells transfected with KOS DNA. This reduction was fully reversed by cotransfection with an ICP0-expressing plasmid. Therefore, ICP0 up-regulates ICP4 expression in the absence of virion-associated proteins during productive infection following transfection of infectious viral DNA. Taken together, these findings demonstrate a role for ICP0 in the induction of E and L gene expression during productive infection and suggest a role for ICP0 in the induction of IE gene expression during the early stage of reactivation from latency which proceeds in the absence of virion-associated proteins.
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页码:2904 / 2915
页数:12
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共 74 条
[11]   ISOLATION AND CHARACTERIZATION OF DELETION MUTANTS OF HERPES-SIMPLEX VIRUS TYPE-1 IN THE GENE ENCODING IMMEDIATE-EARLY REGULATORY PROTEIN-ICP4 [J].
DELUCA, NA ;
MCCARTHY, AM ;
SCHAFFER, PA .
JOURNAL OF VIROLOGY, 1985, 56 (02) :558-570
[12]   ACTIVATION OF IMMEDIATE-EARLY, EARLY, AND LATE PROMOTERS BY TEMPERATURE-SENSITIVE AND WILD-TYPE FORMS OF HERPES-SIMPLEX VIRUS TYPE-1 PROTEIN ICP4 [J].
DELUCA, NA ;
SCHAFFER, PA .
MOLECULAR AND CELLULAR BIOLOGY, 1985, 5 (08) :1997-2008
[13]   RELATIONSHIP BETWEEN POLYADENYLATED AND NONPOLYADENYLATED HERPES-SIMPLEX VIRUS TYPE-1 LATENCY-ASSOCIATED TRANSCRIPTS [J].
DEVIRAO, GB ;
GOODART, SA ;
HECHT, LM ;
ROCHFORD, R ;
RICE, MK ;
WAGNER, EK .
JOURNAL OF VIROLOGY, 1991, 65 (05) :2179-2190
[14]   FINE-STRUCTURE MAPPING AND FUNCTIONAL-ANALYSIS OF TEMPERATURE-SENSITIVE MUTANTS IN THE GENE ENCODING THE HERPES-SIMPLEX VIRUS TYPE-1 IMMEDIATE EARLY PROTEIN VP175 [J].
DIXON, RAF ;
SCHAFFER, PA .
JOURNAL OF VIROLOGY, 1980, 36 (01) :189-203
[15]   IDENTIFICATION OF THE LATENCY-ASSOCIATED TRANSCRIPT PROMOTER BY EXPRESSION OF RABBIT BETA-GLOBIN MESSENGER-RNA IN MOUSE SENSORY NERVE GANGLIA LATENTLY INFECTED WITH A RECOMBINANT HERPES-SIMPLEX VIRUS [J].
DOBSON, AT ;
SEDERATI, F ;
DEVIRAO, G ;
FLANAGAN, WM ;
FARRELL, MJ ;
STEVENS, JG ;
WAGNER, EK ;
FELDMAN, LT .
JOURNAL OF VIROLOGY, 1989, 63 (09) :3844-3851
[16]   AN ANTIGEN ENCODED BY THE LATENCY-ASSOCIATED TRANSCRIPT IN NEURONAL CELL-CULTURES LATENTLY INFECTED WITH HERPES-SIMPLEX VIRUS TYPE-1 [J].
DOERIG, C ;
PIZER, LI ;
WILCOX, CL .
JOURNAL OF VIROLOGY, 1991, 65 (05) :2724-2727