Defective Virions of Herpes Simplex Viruses

Bronson, David L.; Dreesman, Gordon R.; Biswal, Nrusingh C.; Benyésh-Melnick, Matilda

doi:10.1159/000148841

Cited by 78 publications

(37 citation statements)

References 11 publications

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“…We wish to emphasize that minor fragments observed in digests of HSV DNAs cannot be attributed to the presence of defective DNA molecules of high buoyant density of the type reported by Bronson et al (9). The minor fragment patterns were highly reproducible and invariably present in all strains tested even after plaque purification, whereas the heavy density DNA appeared only after serial undiluted passaging and could be easily eliminated.…”

Section: Discussionmentioning

confidence: 58%

Anatomy of herpes simplex virus DNA: strain differences and heterogeneity in the locations of restriction endonuclease cleavage sites.

Hayward¹,

Frenkel²,

Roizman³

1975

Proc. Natl. Acad. Sci. U.S.A.

142

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We are attempting to construct maps of restriction enzyme fragments from the DNA of herpes simplex virus (HSV), and wish to report on some unusual observations relating to the structure of HSV DNA molecules that emerged from these studies. We have also been concerned about possible genetic differences between the various strains and isolates of HSV-1 and HSV-2. Two clinical isolates, designated HSV-1(Fl) and HSV-2(G1), have been used in this laboratory as prototypes of naturally occurring strains and have been passaged at low multiplicity a maximum of four times in HEp-2 (human epidermoid carcinoma) cells to avoid selection of variants in culture. However, established "laboratory" strains characterized by histories of numerous passages outside the human (5), and show apl)roximately 50% homology with 85% matching of base pairs (8). (iv) An additional DNA species with unusually high buoyant density (>1.731 g/cm3) may also be present in viral l)reparations after serial passages at high multiplicity (9). MATERIALS AND METHODSVirus Strains. HSV-1 strains, designated Fl, F5, and F9, andl HSV-2(G1) are independent isolates from patients at the University of Chicago Hospitals (1). HSV-1 (A428) was isolated from human trigeminal ganglion by Dr. A. Nahmias, Atlanta, Ga. All isolates were passaged a maximum of four times at low multiplicity inlHEp-2 cells. Despite these immunological tests, l)r. F. Rapp found in 1973 that the DNA of the Justin strain had a buoyant density pattern similar to that of HSV-2 (personal communication, A. Sabin).We believe that this was probably due to the fact that most of the Justin strain viral D)NA is defective-as much as 80% in some of our preparations. In our laboratory, the Justin strain was identified as HSV-l on the basis of structural polypeptide content and by its D)NA buoyant density and restriction endonuclease cleavage pattern. 1768

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Section: Discussionmentioning

confidence: 58%

Anatomy of herpes simplex virus DNA: strain differences and heterogeneity in the locations of restriction endonuclease cleavage sites.

Hayward¹,

Frenkel²,

Roizman³

1975

Proc. Natl. Acad. Sci. U.S.A.

142

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“…When HSV-1 is serially passaged at a high multiplicity of infection, virus stocks are produced which interfere with the infectivity of standard (non-defective) virus (Bronson et al, 1973;Murray et al, 1975 ;0000-6320 © 1985 (1984) suggested that these two signals may have to be on a contiguous DNA fragment for interference to occur, but the interpretation of their experiments is complicated by the fact that the defective genomes present within a virus stock generally represent a heterogeneous collection of molecules of incompletely defined sequence. In the present studies we have therefore attempted to extend these observations by using plasmids containing cloned copies of functional or inactivated ORIs and the viral 'a' sequence either alone or in combination.…”

Section: Introductionmentioning

confidence: 99%

Mutagenesis of a Herpes Simplex Virus Origin of DNA Replication and its Effect on Viral Interference

Stow

1985

Journal of General Virology

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SUMMARYSupercoiled plasmid molecules containing cloned copies of a DNA fragment which includes a functional herpes simplex virus type 1 origin of DNA replication were cleaved preferentially at two positions within the viral insert by nuclease S1. Plasmids with molecular linker insertions at these sites were constructed, and analysis of two representative plasmids demonstrated the presence of palindromic DNA sequences at the preferred cleavage positions. One of these palindromic sequences occurred within a 90 bp region in which the cis-acting sequences essential for viral origin function had previously been located, Insertion of a linker at this position abolished origin activity, demonstrating an essential role for sequences within the palindrome in the initiation of DNA synthesis. In transfection assays, plasmids containing a functional viral origin of DNA replication markedly interfered with the infectivity of non-defective viral DNA even in the absence of viral encapsidation signals. Inactivation of the origin greatly reduced this effect on DNA infectivity, suggesting that viral interference may be mediated by a mechanism involving the replicative machinery.

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“…Whereas recently plaque-purified virus stocks predominantly contain standard HSV genomes, defective virus populations obtained through serial undiluted propagation of such stocks have been shown to contain variable proportions of variant DNA molecules in which greater than 90% of the parental viral DNA sequences have been replaced by multiple head-totail reiterations of sequences derived from defined locations of the parental genome (9)(10)(11)(12)(13)(14)(15). On the basis of the origin of DNA sequences contained within their repeat units, defective HSV genomes have been divided into two classes.…”

mentioning

confidence: 99%

Replication of herpes simplex virus DNA: localization of replication recognition signals within defective virus genomes.

Vlazny¹,

Frenkel²

1981

Proc. Natl. Acad. Sci. U.S.A.

136

102

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Serially passaged herpes simplex virus type 1 (HSV-1) strain Justin was previously shown to contain defective virus genomes consisting ofhead-to-tail reiterations ofsequences derived from the end ofthe S component ofthe standard virus DNA. Cotransfection of purified monomeric defective genome repeat units with foster helper virus DNAs onto rabbit skin cells resulted in regeneration and replication of concatemeric defective DNA molecules which were successfully encapsidated. Thus, defective HSV-1 (Justin) genomes contain, within their limited DNA sequences, a sufficient set of recognition sites required for HSV DNA replication and packaging. The arrangement of repeat units within the regenerated defective virus genomes was consistent with their replication by a rolling circle mechanism in which a single repeat unit served as the circularized template. This replication occurred most actively late after infection and could be shown to be inhibited by low concentrations of phosphonoacetate known to inhibit the HSV-specified viral DNA polymerase selectively. The resultant concatemers were shown to be cleaved to M 100 X 106 DNA molecules which were terminated at one end with the proper ac end sequence of the parental standard virus DNA.The standard genome of herpes simplex virus type 1 (HSV-1) is a linear, double-stranded DNA molecule with Mr 100 X 106 (1, 2). Topologically, the standard HSV DNA is terminally reiterated and consists oftwo covalently linked components, L and S, each of which contains a stretch of unique viral DNA sequences surrounded by inverted repeats: the unique sequences of L (UL) surrounded by the ab and b'a' inverted repeat sequences, and the unique sequences ofS (Us) bounded by the ac and c'a' inverted repeats (3-6). The L and S components have been shown to invert relative to each other, yielding equimolar amounts of four isomeric structures. (3,(5)(6)(7)(8).Whereas recently plaque-purified virus stocks predominantly contain standard HSV genomes, defective virus populations obtained through serial undiluted propagation of such stocks have been shown to contain variable proportions of variant DNA molecules in which greater than 90% of

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Defective Virions of Herpes Simplex Viruses

Cited by 78 publications

References 11 publications

Anatomy of herpes simplex virus DNA: strain differences and heterogeneity in the locations of restriction endonuclease cleavage sites.

Anatomy of herpes simplex virus DNA: strain differences and heterogeneity in the locations of restriction endonuclease cleavage sites.

Mutagenesis of a Herpes Simplex Virus Origin of DNA Replication and its Effect on Viral Interference

Replication of herpes simplex virus DNA: localization of replication recognition signals within defective virus genomes.

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