Herpes simplex virus type 1 DNA cleavage and encapsidation require the product of the UL28 gene: isolation and characterization of two UL28 deletion mutants

Tengelsen, Leslie; Pederson, Nels E.; Shaver, Patti R.; Wathen, Michael W.; Homa, Fred L.

doi:10.1128/jvi.67.6.3470-3480.1993

Cited by 119 publications

(75 citation statements)

References 50 publications

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“…Previously described procedures were used for growth and maintenance of African green monkey kidney cells (Vero; ATCC CCL 81 [26]). Vero-derived C1 (49), F3 (19), D6 (11), and 3A6 cells were grown in Dulbecco modified Eagle medium plus 10% fetal bovine serum and 0.5 mg of G418 (Geneticin; GIBCO-BRL) per ml. The KOS strain of HSV-1 was used as the wild-type strain.…”

Section: Methodsmentioning

confidence: 99%

“…The KOS strain of HSV-1 was used as the wild-type strain. The HSV-1 UL28 deletion mutant GCB (49) was grown on UL28-complementing cell line C1. The UL26 and UL27 null viruses, KUL26⌬Z (19) and K082 (11), were grown on 3A6 and D6 cells, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Empty A capsids are thought to result from abortive attempts at DNA encapsidation (39). In studies with HSV-1 mutants, the UL6, UL15, UL25, UL28, UL32, and UL33 genes have been shown to be required for processing and packaging of viral DNA (1,2,4,6,28,29,37,40,44,45,49,55,56). In addition, the product of the UL12 gene has been shown to play an important but not essential role in cleavage and packaging.…”

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confidence: 99%

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The Product of the Herpes Simplex Virus Type 1 UL25 Gene Is Required for Encapsidation but Not for Cleavage of Replicated Viral DNA

McNab

Desai

Person

et al. 1998

J Virol

163

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The herpes simplex virus type 1 (HSV-1) UL25 gene contains a 580-amino-acid open reading frame that codes for an essential protein. Previous studies have shown that the UL25 gene product is a virion component (M. A. Ali et al., Virology 216:278-283, 1996) involved in virus penetration and capsid assembly (C. Addison et al., Virology 138:246-259, 1984). In this study, we describe the isolation of a UL25 mutant (KUL25NS) that was constructed by insertion of an in-frame stop codon in the UL25 open reading frame and propagated on a complementing cell line. Although the mutant was capable of synthesis of viral DNA, it did not form plaques or produce infectious virus in noncomplementing cells. Antibodies specific for the UL25 protein were used to demonstrate that KUL25NS-infected Vero cells did not express the UL25 protein. Western immunoblotting showed that the UL25 protein was associated with purified, wild-type HSV A, B, and C capsids. Transmission electron microscopy indicated that the nucleus of Vero cells infected with KUL25NS contained large numbers of both A and B capsids but no C capsids. Analysis of infected cells by sucrose gradient sedimentation analysis confirmed that the ratio of A to B capsids was elevated in KUL25NS-infected Vero cells. Following restrictionenzyme digestion, specific terminal fragments were observed in DNA isolated from KUL25NS-infected Vero cells, indicating that the UL25 gene was not required for cleavage of replicated viral DNA. The latter result was confirmed by pulsed-field gel electrophoresis (PFGE), which showed the presence of genome-size viral DNA in KUL25NS-infected Vero cells. DNase I treatment prior to PFGE demonstrated that monomeric HSV DNA was not packaged in the absence of the UL25 protein. Our results indicate that the product of the UL25 gene is required for packaging but not cleavage of replicated viral DNA.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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The Product of the Herpes Simplex Virus Type 1 UL25 Gene Is Required for Encapsidation but Not for Cleavage of Replicated Viral DNA

McNab

Desai

Person

et al. 1998

J Virol

163

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“…The translocation of concatenated viral DNA into procapsids and its cleavage at packaging sites is not understood. Recent studies with herpes simplex virus type 1 (HSV-1) mutants defective in UL6, UL15, UL25, UL28, UL32 or UL33 suggest that these genes are essentially involved in viral DNA cleavage and packaging, since cells infected with these mutants produce only B capsids lacking DNA (Al-Kobaisi et al 1991;Baines et al 1997;Weller 1996, 1998;McNab et al 1998;Patel et al 1996;Tengelsen et al 1993;Yu et al 1997). The respective homologues of these genes in HCMV are UL104, UL89, UL77, UL56, UL52 and UL51 (Chee et al 1990).…”

Section: Terminase Inhibitorsmentioning

confidence: 99%

Other Inhibitors of Viral Enzymes and Functions

Zimmermann

Hewlett²,

Rübsamen‐Waigmann

2009

Antiviral Strategies

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Until the end of the 1970s, the mainstays of antiviral chemotherapy were nucleoside analogues that targeted virus polymerase, in particular, the herpesvirus DNA polymerase. The scourge of HIV triggered an unprecedented commitment to identify novel antivirals, and these efforts transformed antiviral therapy into the modern, sophisticated treatment form described in this book, with targets such as the reverse transcriptase and the protease as well as the entry of the human immunodeficiency virus. As the regulation of human pathogenic virus growth cycles became more understandable, the realisation grew that these pathogens had more than one Achilles heel that might be suitable targets for small molecules with antiviral activity. This chapter addresses those "other" targets as well as other approaches to the tried and tested polymerase inhibitors, the so-called non-nucleoside inhibitors of reverse transcriptase.

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“…Most terminases are hetero-oligomers with each subunit carrying a di¡erent function [11^16]. Mutations in any of the encoding genes lead to an accumulation of empty procapsids/proheads and uncleaved concatemeric DNA in the nucleus [9,17,18]. Our previous data together with the reports from Krosky et al [19] and Underwood et al [20] indicated that the terminase of HCMV consists of the highly conserved proteins pUL56 and pUL89 [212 6].…”

Section: Introductionmentioning

confidence: 99%

Insights into the structure of human cytomegalovirus large terminase subunit pUL56

Savva

Holzenburg

Bogner³

2004

FEBS Letters

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Terminases are a class of proteins which catalyze the generation of unit-length genomes during DNA packaging. These essential proteins are conserved throughout the herpesviruses and many double-stranded DNA bacteriophages. We have determined the structure of the large terminase subunit pUL56 of human cytomegalovirus, a highly pathogenic virus, to 2.6 nm resolution. Image analysis of puri¢ed pUL56 suggests that the molecule exists as a dimer formed by the association of two ring-like structures positioned on top of each other and connected by a pronounced density on one side. The 3D reconstruction of pUL56 provides ¢rst structural insights into the active protein.

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Herpes simplex virus type 1 DNA cleavage and encapsidation require the product of the UL28 gene: isolation and characterization of two UL28 deletion mutants

Cited by 119 publications

References 50 publications

The Product of the Herpes Simplex Virus Type 1 UL25 Gene Is Required for Encapsidation but Not for Cleavage of Replicated Viral DNA

The Product of the Herpes Simplex Virus Type 1 UL25 Gene Is Required for Encapsidation but Not for Cleavage of Replicated Viral DNA

Other Inhibitors of Viral Enzymes and Functions

Insights into the structure of human cytomegalovirus large terminase subunit pUL56

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