Herpes simplex virus mRNA species mapping in EcoRI fragment I

Hall, Lucinda M. C.; Draper, Ken; Frink, R J; Costa, Robert H.; Wagner, Edward K.

doi:10.1128/jvi.43.2.594-607.1982

Cited by 93 publications

(80 citation statements)

References 20 publications

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“…The gD coding sequence (394 codons) terminates approximately 100 bp upstream of the 1.6 kb early MM 5' terminus. The presence of families of 3' co-terminal mIm's has been previously described for other regicns of the HSV-1 genane (37,38,39). In sane instances, the snaller members of the family have been shcon to specify proteins fran coding sequences present in the 3' untranslated region of the larger members (37,40).…”

Section: Discussionmentioning

confidence: 81%

Characterization of the herpes simplex virus type 1 glycoprotein D mRNA and expression of this protein in Xenopus oocytes

Watson¹,

Colberg-Poley²,

Marcus-Sekura³

et al. 1983

Nucleic Acids Research

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We have identified and characterized a 3.0 kilobase (kb) mRNA containing coding sequences of the herpes simplex virus type 1 (HSV-1) glycoprotein D (gD) gene. The synthesis of this 3.0 kb mRNA was unaffected by the presence of cytosine arabinoside, but was made in greatly reduced amounts in cells infected with HSV-1 in the presence of cycloheximide: it was, therefore, classified as an early mRNA. By nuclease protection experiments, it was found that the 3.0 kb mRNA is unspliced and, further, that it is 3' co-terminal with a smaller 1.6 kb early mRNA which is transcribed from a DNA sequence 3' to the gD coding sequence. We describe the use of the Xenopus laevis oocyte system to produce HSV-1 gD in vitro. Oocytes injected with mRNA isolated from HSV-1-infected Vero cells synthesized gD, which was identified by immunoprecipitation. Injection of a plasmid clone containing the HSV-1 BamHI J fragment (0.89 to 0.93 map units) into the nuclei of Xenopus oocytes also resulted in synthesis of gD.

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Section: Discussionmentioning

confidence: 81%

Characterization of the herpes simplex virus type 1 glycoprotein D mRNA and expression of this protein in Xenopus oocytes

Watson¹,

Colberg-Poley²,

Marcus-Sekura³

et al. 1983

Nucleic Acids Research

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“…Our finding that the overrepresentation of 92/91K (VP11/ 12) in PREPs is 2.45-fold, compared with that in L-particles, argues for nonrandom incorporation of at least some tegument proteins during assembly of PREPs, because the early/late temporal classification of the UL46 gene (8,28) which encodes these proteins suggests that they should be present in reduced amounts in WT virus-infected cells treated with an inhibitor of viral DNA synthesis or in cells infected with the ambUL8 virus. Zhang and McKnight (28) have suggested that essential tegument proteins, such as VP16, provide a fixed framework around which nonessential proteins assemble.…”

Section: Discussionmentioning

confidence: 78%

PREPs: herpes simplex virus type 1-specific particles produced by infected cells when viral DNA replication is blocked

Dargan

Patel

Subak‐Sharpe

1995

J Virol

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Herpes simplex virus (HSV)-infected cells produce not only infectious nucleocapsid-containing virions butalso virion-related noninfectious light particles (L-particles) composed of the envelope and tegument components of the virus particle (J. F. Szilágyi and C. Cunningham, J. Gen. Virol. 62:661-668, 1991). We show that BHK and MeWO cells infected either with wild-type (WT) HSV type 1 (HSV-1) in the presence of viral DNA replication inhibitors (cytosine-␤-D-arabinofuranoside, phosphonoacetic acid, and acycloguanosine) or with a viral DNA replication-defective mutant of HSV-1 (ambUL8) synthesize a new type of virus-related particle that is morphologically similar to an L-particle but differs in its relative protein composition. These novel particles we term pre-viral DNA replication enveloped particles (PREPs). The numbers of PREPs released into the culture medium were of the same order as those of L-particles from control cultures. The particle/PFU ratios of different PREP stocks ranged from 6 ؋ 10 5 to 3.8 ؋ 10 8 , compared with ratios of 3 ؋ 10 3 to 1 ؋ 10 4 for WT L-particle stocks. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western immunoblot analyses revealed that true late proteins, such as 273K (VP1-2), 82/81K (VP13/14), and gC (VP8), were greatly reduced or absent in PREPs and that gD (VP17) and 40K proteins were also underrepresented. In contrast, the amounts of proteins 175K (VP4; IE3), 92/91K (VP11/12), 38K (VP22), and gE (with BHK cells) were increased. The actual protein composition of PREPs showed some cell line-dependent differences, particularly in the amount of gE. PREPs were biologically competent and delivered functional Vmw65 (VP16; ␣TIF) to target cells, but the efficiency of complementation of the HSV-1 (strain 17) mutant in1814 was 10 to 30% of that of WT L-particles.

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“…Alternatively, if coordinate transcription of UL46 and UL47 occurs, the stoichiometry of these proteins relative to each other could be important. Since the transient-expression assays, unlike the current experiments, did not transcriptionally separate UL46 and UL47, but rather interrupted protein expression through the insertion of translational stop codons, the effect of putative coordinate transcription of these 3' coterminal mRNAs would not have been measured (15,34).…”

Section: Discussionmentioning

confidence: 85%

Role of herpes simplex virus type 1 UL46 and UL47 in alpha TIF-mediated transcriptional induction: characterization of three viral deletion mutants

Zhang

Sirko

Mcknight

1991

J Virol

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The transcriptional induction of the a or immediate-early gene class of herpes simplex virus type 1 effected by the a trans-induction factor (aTIF, ICP25, VP16, Vmw65) requires an a-specific cis-acting site. Increased transcription does not result from the direct, independent binding of aTIF, but rather from an aTIFdependent formation of a protein-DNA complex containing, in addition to aTIF, at least one host cell factor. One of the host factors is a POU domain protein which recognizes an octamer element in the a-specific consensus. There is evidence that aTIF may drive the formation of multiple protein-DNA complexes containing a POU protein and additional host factors. Previously, the gene products of UL46 and UL47 have been implicated in modulating the aTIF-dependent transcriptional induction of a genes. Our current studies have extended these analyses from a transient-expression system to a series of viral deletion mutants. In these studies we demonstrate that neither UL46nor UL47-encoded gene product, either separately or in combination, is required for viral growth in cell culture. The absence of UL47 reduces by up to 80% the ability of the virus to induce an a-regulated thymidine kinase reporter gene resident in 143TK-cells. Autoradiograms of [35S]methionine pulse-labeled infected cell proteins, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, show that deleting UL46 and/or UL47 has no discernable effect on the synthesis of aTIF or aTIF-containing proteins. Subsequent Western immunoblot analysis, with rabbit anti-aTIF antibodies made to an otTIF-Staphylococcus aureus protein A fusion, demonstrated that the accumulation and steady-state levels of aTIF or aTIF-containing proteins was indistinguishable from that of the thymidine kinase-negative isogenic parental virus, RA305. * Corresponding author. able to complex with aTIF (3,12,21,23,24,32,37,44,49). Complex formation appears to be mediated through the POU domain present in these proteins, which recognizes an octamer element contained within the a element (gyATGN TAATgarattcyttgnggg) (3,12,18,21,24,36). A recent report suggests that aTIF may drive the formation of multiple protein-DNA complexes containing a POU protein and additional host factors, one of which may be cell type specific (21).Previous work has identified a role for at least two additional viral factors, encoded by HSV-1 UL46 and UL47, in aTIF-dependent transcriptional induction (33,34). In transient-expression assays the gene product of UL46 was able to enhance aTIF-mediated induction of an aTK reporter construct 2to 3-fold, while the gene product of UL47 acted to reduce the levels of aTIF-mediated induction 2.5to 3-fold in the presence or absence of UL46. These results were the first report of additional viral factors involved in a-gene induction, and they predicted the presence of a regulatory gene cluster composed of UL46, UL47, and aTIF (UL48) (33,34).The goals of our current study were to investigate the roles of UL46 and UL47 in the context of the virus to determi...

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Herpes simplex virus mRNA species mapping in EcoRI fragment I

Cited by 93 publications

References 20 publications

Characterization of the herpes simplex virus type 1 glycoprotein D mRNA and expression of this protein in Xenopus oocytes

Characterization of the herpes simplex virus type 1 glycoprotein D mRNA and expression of this protein in Xenopus oocytes

PREPs: herpes simplex virus type 1-specific particles produced by infected cells when viral DNA replication is blocked

Role of herpes simplex virus type 1 UL46 and UL47 in alpha TIF-mediated transcriptional induction: characterization of three viral deletion mutants

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