Unity and Diversity in the Herpesviruses

SUMMARYThe addition of TPA (phorbol-12-myristate-13-acetate) to cultures during the lytic infection with herpesvirus saimiri led to an enhanced and accelerated production of polypeptides induced by H. saimiri and to a rapid shut-down of host cell protein synthesis and allowed a detailed analysis of the protein patterns. Analysis of sequential protein synthesis in owl monkey kidney cells lytically infected with H. saimiri 11 permitted the identification of 31 virus-induced polypeptides. The use of the amino acid analogues canavanine (for arginine) and azetidine (for proline) in parallel allowed experiments on the identification of proteins synthesized early and late during lytic infection.

Section: Dependence Of Virus-induced Protein Synthesis On Dna Replicamentioning

confidence: 47%

Section: Introductionmentioning

confidence: 99%

Herpesvirus Saimiri-induced Proteins in Lytically Infected Cells. I. Time-ordered Synthesis

Modrow¹,

Wolf²

1983

“…150K and other components of the capsid structure resemble those of a number of other herpesviruses (R. W. Honess et al, unpublished results). HVS also specifies a thymidine kinase and a DNA polymerase (O'Hare, 1981), but beyond these rudimentary group characteristics (Honess & Watson, 1977) the gene products of HVS bear some more striking resemblances to those of EBV. For example, the 160K polypeptide of HVS, a poorly or nonglycosylated component of the virus envelope (Randall & Honess, 1980, has a counterpart in the enveloped particle of EBV (Dolyniuk et al, 1979 a, b) and the poorly glycosylated 160K to 170K component of the EBV membrane antigen (MA; Qualtiere & Pearson, 1979;ThorleyLawson & Edson, 1979;North et al, 1980).…”

Section: Demonstration O F a N Electrophoretically Distinct Nuclear Pmentioning

confidence: 99%

“…of 71.6 x 106 and a mean composition of36~ G + C bracketed by multiple copies of a 0.83 × 106 sequence with a mean composition of 71 ~o G + C. These repeated sequences are in head-to-tail, contiguous arrays and are oriented in the same direction at each end of the genome, with a total of about 30 repeat units per genome distributed randomly between the two molecular termini (Bornkamm et al, 1976;Fleckenstein et al, 1975Fleckenstein et al, , 1978Fleckenstein, 1979). This general arrangement of unique and reiterated sequences constitutes a distinct subgroup within the herpesviruses and the mean nucleotide composition of the unique sequences is among the lowest observed for any member of this virus group (Honess & Watson, 1977). However, there is little published information on the nature, location or control of the genes encoded by this herpesvirus.…”

Section: Introductionmentioning

confidence: 99%

Proteins Specified by Herpesvirus Saimiri: Identification and Properties of Virus-specific Polypeptides in Productively Infected Cells

Randall¹,

Honess²,

O’Hare³

1983

Self Cite

SUMMARYThe synthesis and accumulation of more than 30 virus-induced polypeptides was detected after infection of permissive primate cells with high multiplicities of herpesvirus saimiri. These virus-induced polypeptides had apparent mol. wt. of from 12000 (12K) to 250K and differed in molar abundance by up to two orders of magnitude. The majority of virus-induced polypeptides satisfied multiple criteria for their virus specificity. Polypeptides of ll0K, 76K, 51K and 31K to 29K were synthesized at maximum rates early in infection, but the majority of proteins were made at high rates late in infection. Virus-specific polypeptides were substrates for a number of post-synthetic modifications. The l17K, 85K, 76K, 31K and 30K polypeptides were each processed to forms with altered electrophoretic mobility. The 117K and 85K polypeptides were among the virus-specified substrates for glycosylation and virus-induced polypeptides of 59K, 51K, 30K and 26K were phosphorylated. The early 51K phosphoprotein and the 30K to 31K polypeptides were rapidly translocated to the nucleus of infected cells. The 31K polypeptide was processed to a 29K product which remained stably associated with the nuclear fraction. The 30K nuclear protein was shown to be the precursor of a 28K polypeptide which was released in a soluble form into the cytoplasmic fraction and the culture medium of cells at late times in the virus growth cycle. Many other polypeptides accumulated slowly in nuclear (e.g. 250K, 150K, 130K, 110K and 38K) or in cytoplasmic (e.g. 117K, 85K and 28K) fractions Of infected cells in tbrms which could be differentiated by the use of detergents or differences in stability to salt extraction. The mol. wt., relative molarities and some features of the post-synthetic processing of herpesvirus saimiri polypeptides more closely resembled the properties of gene products of Epstein-Barr virus than those of herpes simplex virus.

“…Herpesviruses have been divided into three subgroups, alpha-, beta-and gammaherpesviruses, based on differences in their biological properties and some aspects of their molecular biology (Honess & Watson, 1977;Roizman, 1982;Honess, 1984). Typical alphaherpesviruses, such as herpes simplex virus type 1 (HSV-1), have relatively broad in vitro host ranges and short cycles of virus replication in permissively infected cells.…”

Section: Introductionmentioning

confidence: 99%

Asynchronous Expression of the Immediate-Early Protein of Herpesvirus Saimiri in Populations of Productively Infected Cells

Randall¹,

Newman²,

Honess³

1985

Self Cite

SUMMARYThe time course of virus replication in cultures of permissive cells infected with high multiplicities of herpesvirus saimiri (HVS), a gammaherpesvirus, is protracted relative to the replication of herpes simplex virus (HSV), an alphaherpesvirus, under similar conditions. The basis for this difference was investigated by quantitative immunofluorescence microscopy exploiting monoclonal antibodies specific to the HVS 52000 mol. wt. immediate-early polypeptide (IE 52K) and to delayed-early (DE 51K, DE 110K) and late (130K and capsid proteins) gene products to measure the timing of gene expression in individual cells of infected cultures. The timing of the transitions from IE to DE and from DE to late protein synthesis occurred at proportionately different intervals in the growth cycle of HVS, relative to that of HSV. In particular, the DE to late transition occurred relatively later in HVS infections. However, asynchrony in the events leading to the expression of the first class of virus proteins (IE 52K) was the main source of the extended course of HVS replication in populations of infected cells. This asynchrony was not modified significantly by infection at different stages of the host cell-cycle and was reduced, but not overcome, by very high applied multiplicities of infection. Double-antibody staining revealed a positive correlation between the accumulation of high concentrations of parental virus particles at perinuclear sites and early detection of HVS IE 52K gene expression. Both of these events remained sensitive to a microtubule poison (colcemid) for many hours after infection with HVS, whereas the rapid and synchronous expression of the IE 175K protein (ICP4) in HSV-1-infected cells was insensitive to post-infection exposures to this drug. We conclude that significant differences in early stages of virus entry and intracellular processing which precede immediate-early gene expression are largely responsible for differences between the replicative cycles of these representatives of gamma-and alphaherpesviruses in cultures of permissive cells.