Anatomy of herpes simplex virus DNA. III. Characterization of defective DNA molecules and biological properties of virus populations containing them

N. Frenkel, R. J. Jacob, R. W. Honess, Gary Selwyn Hayward, H. Locker, B. Roizman

Research output: Contribution to journalArticle

Abstract

We have characterized the virus progeny and its DNA from plaque purified and undiluted passages of herpes simplex virus 1 in HEp-2 cells. Specifically, (i) infectious virus yields declined progressively in passages 1 through 10 and gradually increased at passages 11 through 14. The yields correlated with PFU/particle ratios. (ii) In cells infected with virus from passages 6 through 10, there was an overproduction of an early viral polypeptide (no.4) and a delay in the synthesis of late viral proteins. In addition, the virus in these passages interfered with the replication of a nondefective marker virus. Cells infected with passage 14 virus produced normal amounts of polypeptide 4 and, moreover, this virus showed minimal interfering capacity. (iii) In addition to DNA of density 1.726 g/cm 3, which was the sole component present in viral progeny of passage O, passages 6 through 14 contained one additional species (rho 1.732) and in some instances (passages 6 and 10) also DNA of an intermediate buoyant density. The ratio of rho 1.732 to rho 1.726 DNA increased to a maximum of 4 in passages 6 through 9 and gradually decreased to 1 in passages 10 through 14. (iv) 1.732 DNA cannot be differentiated from rho 1.726 DNA with respect to size; however, it has no Hin III restriction enzyme cleavage sites and yields only predominantly two kinds of fragments with molecular weights of 5.1x10 6 and 5.4x10 6 upon digestion with EcoRI enzyme. (v) Partial denaturation profiles of purified rho 1.732 DNA from passage 14 revealed the presence of two types of tandemly repeated units corresponding roughly in size to the EcoRI fragments and situated in different molecules. (vi) In addition to the two kinds of rho 1.732 molecules consisting of tandem repeat units of different sizes, other evidence for the diversity of defective DNA molecules emerged from comparisons of specific infectivity and interfering capacity of the progeny from various passages. The data suggest that some of the particles with DNA of normal buoyant density (1.726) must also be defective since the capacity to interfere and to produce an excess of polypeptide 4 did not appear to be proportional to the amount of high buoyant density defective DNA. The data suggest that defective interfering particles are replaced by defective particles with diminished capacity to interfere and that more than one species of defective DNA molecules evolves on serial preparation of HSV.

Original languageEnglish (US)
Pages (from-to)153-167
Number of pages15
JournalJournal of Virology
Volume16
Issue number1
StatePublished - 1975
Externally publishedYes

Fingerprint

herpes simplex
DNA viruses
Simplexvirus
Anatomy
Viruses
viruses
DNA
Population
polypeptides
Peptides
Defective Viruses
Human herpesvirus 1
Tandem Repeat Sequences
tandem repeat sequences
viral proteins
Human Herpesvirus 1
Viral Proteins
cells
Enzymes
enzymes

ASJC Scopus subject areas

  • Immunology

Cite this

Anatomy of herpes simplex virus DNA. III. Characterization of defective DNA molecules and biological properties of virus populations containing them. / Frenkel, N.; Jacob, R. J.; Honess, R. W.; Hayward, Gary Selwyn; Locker, H.; Roizman, B.

In: Journal of Virology, Vol. 16, No. 1, 1975, p. 153-167.

Research output: Contribution to journalArticle

@article{892961b39485474591b9cbebe859aba9,
title = "Anatomy of herpes simplex virus DNA. III. Characterization of defective DNA molecules and biological properties of virus populations containing them",
abstract = "We have characterized the virus progeny and its DNA from plaque purified and undiluted passages of herpes simplex virus 1 in HEp-2 cells. Specifically, (i) infectious virus yields declined progressively in passages 1 through 10 and gradually increased at passages 11 through 14. The yields correlated with PFU/particle ratios. (ii) In cells infected with virus from passages 6 through 10, there was an overproduction of an early viral polypeptide (no.4) and a delay in the synthesis of late viral proteins. In addition, the virus in these passages interfered with the replication of a nondefective marker virus. Cells infected with passage 14 virus produced normal amounts of polypeptide 4 and, moreover, this virus showed minimal interfering capacity. (iii) In addition to DNA of density 1.726 g/cm 3, which was the sole component present in viral progeny of passage O, passages 6 through 14 contained one additional species (rho 1.732) and in some instances (passages 6 and 10) also DNA of an intermediate buoyant density. The ratio of rho 1.732 to rho 1.726 DNA increased to a maximum of 4 in passages 6 through 9 and gradually decreased to 1 in passages 10 through 14. (iv) 1.732 DNA cannot be differentiated from rho 1.726 DNA with respect to size; however, it has no Hin III restriction enzyme cleavage sites and yields only predominantly two kinds of fragments with molecular weights of 5.1x10 6 and 5.4x10 6 upon digestion with EcoRI enzyme. (v) Partial denaturation profiles of purified rho 1.732 DNA from passage 14 revealed the presence of two types of tandemly repeated units corresponding roughly in size to the EcoRI fragments and situated in different molecules. (vi) In addition to the two kinds of rho 1.732 molecules consisting of tandem repeat units of different sizes, other evidence for the diversity of defective DNA molecules emerged from comparisons of specific infectivity and interfering capacity of the progeny from various passages. The data suggest that some of the particles with DNA of normal buoyant density (1.726) must also be defective since the capacity to interfere and to produce an excess of polypeptide 4 did not appear to be proportional to the amount of high buoyant density defective DNA. The data suggest that defective interfering particles are replaced by defective particles with diminished capacity to interfere and that more than one species of defective DNA molecules evolves on serial preparation of HSV.",
author = "N. Frenkel and Jacob, {R. J.} and Honess, {R. W.} and Hayward, {Gary Selwyn} and H. Locker and B. Roizman",
year = "1975",
language = "English (US)",
volume = "16",
pages = "153--167",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "1",

}

TY - JOUR

T1 - Anatomy of herpes simplex virus DNA. III. Characterization of defective DNA molecules and biological properties of virus populations containing them

AU - Frenkel, N.

AU - Jacob, R. J.

AU - Honess, R. W.

AU - Hayward, Gary Selwyn

AU - Locker, H.

AU - Roizman, B.

PY - 1975

Y1 - 1975

N2 - We have characterized the virus progeny and its DNA from plaque purified and undiluted passages of herpes simplex virus 1 in HEp-2 cells. Specifically, (i) infectious virus yields declined progressively in passages 1 through 10 and gradually increased at passages 11 through 14. The yields correlated with PFU/particle ratios. (ii) In cells infected with virus from passages 6 through 10, there was an overproduction of an early viral polypeptide (no.4) and a delay in the synthesis of late viral proteins. In addition, the virus in these passages interfered with the replication of a nondefective marker virus. Cells infected with passage 14 virus produced normal amounts of polypeptide 4 and, moreover, this virus showed minimal interfering capacity. (iii) In addition to DNA of density 1.726 g/cm 3, which was the sole component present in viral progeny of passage O, passages 6 through 14 contained one additional species (rho 1.732) and in some instances (passages 6 and 10) also DNA of an intermediate buoyant density. The ratio of rho 1.732 to rho 1.726 DNA increased to a maximum of 4 in passages 6 through 9 and gradually decreased to 1 in passages 10 through 14. (iv) 1.732 DNA cannot be differentiated from rho 1.726 DNA with respect to size; however, it has no Hin III restriction enzyme cleavage sites and yields only predominantly two kinds of fragments with molecular weights of 5.1x10 6 and 5.4x10 6 upon digestion with EcoRI enzyme. (v) Partial denaturation profiles of purified rho 1.732 DNA from passage 14 revealed the presence of two types of tandemly repeated units corresponding roughly in size to the EcoRI fragments and situated in different molecules. (vi) In addition to the two kinds of rho 1.732 molecules consisting of tandem repeat units of different sizes, other evidence for the diversity of defective DNA molecules emerged from comparisons of specific infectivity and interfering capacity of the progeny from various passages. The data suggest that some of the particles with DNA of normal buoyant density (1.726) must also be defective since the capacity to interfere and to produce an excess of polypeptide 4 did not appear to be proportional to the amount of high buoyant density defective DNA. The data suggest that defective interfering particles are replaced by defective particles with diminished capacity to interfere and that more than one species of defective DNA molecules evolves on serial preparation of HSV.

AB - We have characterized the virus progeny and its DNA from plaque purified and undiluted passages of herpes simplex virus 1 in HEp-2 cells. Specifically, (i) infectious virus yields declined progressively in passages 1 through 10 and gradually increased at passages 11 through 14. The yields correlated with PFU/particle ratios. (ii) In cells infected with virus from passages 6 through 10, there was an overproduction of an early viral polypeptide (no.4) and a delay in the synthesis of late viral proteins. In addition, the virus in these passages interfered with the replication of a nondefective marker virus. Cells infected with passage 14 virus produced normal amounts of polypeptide 4 and, moreover, this virus showed minimal interfering capacity. (iii) In addition to DNA of density 1.726 g/cm 3, which was the sole component present in viral progeny of passage O, passages 6 through 14 contained one additional species (rho 1.732) and in some instances (passages 6 and 10) also DNA of an intermediate buoyant density. The ratio of rho 1.732 to rho 1.726 DNA increased to a maximum of 4 in passages 6 through 9 and gradually decreased to 1 in passages 10 through 14. (iv) 1.732 DNA cannot be differentiated from rho 1.726 DNA with respect to size; however, it has no Hin III restriction enzyme cleavage sites and yields only predominantly two kinds of fragments with molecular weights of 5.1x10 6 and 5.4x10 6 upon digestion with EcoRI enzyme. (v) Partial denaturation profiles of purified rho 1.732 DNA from passage 14 revealed the presence of two types of tandemly repeated units corresponding roughly in size to the EcoRI fragments and situated in different molecules. (vi) In addition to the two kinds of rho 1.732 molecules consisting of tandem repeat units of different sizes, other evidence for the diversity of defective DNA molecules emerged from comparisons of specific infectivity and interfering capacity of the progeny from various passages. The data suggest that some of the particles with DNA of normal buoyant density (1.726) must also be defective since the capacity to interfere and to produce an excess of polypeptide 4 did not appear to be proportional to the amount of high buoyant density defective DNA. The data suggest that defective interfering particles are replaced by defective particles with diminished capacity to interfere and that more than one species of defective DNA molecules evolves on serial preparation of HSV.

UR - http://www.scopus.com/inward/record.url?scp=0016707164&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0016707164&partnerID=8YFLogxK

M3 - Article

C2 - 166202

AN - SCOPUS:0016707164

VL - 16

SP - 153

EP - 167

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 1

ER -