Observations on the Fine Structure of Type 5 Adenovirus

Epstein, M. A.

doi:10.1083/jcb.6.3.523

Cited by 21 publications

(6 citation statements)

References 12 publications

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“…That DNase was able to act directly on the nucleoid of herpes virus without some preliminary pretreatment of the particle was in marked contrast to earlier findings with the adenovirus, an agent without an outer limiting membrane (34); in that case the nucleoid was only digested by enzyme when the protein of the viral outer coat had first been denatured with mild alkali (4). On the other hand, the nucleoid of the Rous virus which has an outer limiting membrane (28) like that of herpes, was susceptible to RNase digestion immediately after fixation (2).…”

Section: Discussioncontrasting

confidence: 57%

Observations on the Fine Structure of Mature Herpes Simplex Virus and on the Composition of Its Nucleoid

Epstein

1962

The Journal of Experimental Medicine

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P y~s 1 To 5 (Received for publication, June 12, 1961) In recent work with adenovirus and the Rous virus, specific enzymic digestions were applied to samples of purified pelleted particles, and the resulting changes caused within the particles were then observed in thin sections with the electron microscope; in this way the site and nature of viral nucleic acid were determined (1-5). Since the accuracy of the findings was confirmed in each case by concomitant specific staining of the purified material for nucleic acids (2, 4), it was considered that the results of applying enzyme digestions and electron microscopy alone to virus particles unseparated from cells might safely be relied upon, such preparations being, of course, unsuitable for the additional checking of observations cytochemically.An approach of this kind seemed particularly appropriate in relation to herpes virus, for the mature particles can be readily recognized and their tendency to accumulate at the cell surface is well known (6-9). Furthermore, although cytochemical investigations of the nuclear inclusions arising in herpes infections have demonstrated that they contain much deoxyribonucleic acid (DNA) (10) little is known about the composition of the virus. It has sometimes been assumed that the latter also includes nucleic acid of this type (11), but even though excess DNA is in fact synthesized by cells very soon after infection with herpes virus (12), no connection has been established between this material and the constituents of the agent itself.Experiments were accordingly undertaken in which herpes-infected HeLa cells were examined in thin sections with the electron microscope so that the mature virus particles around them might be studied after treatment with various fi afives, specific nucleases, and in different embedding media, both with and without electron staining. Tests for the biological activity of the cultures examined were included in the first part of the work. The results obtained in this investigation are now reported. Materials and MethodsVirus Strain.--The HFEM strain of herpes virus has been used; it was received frozen at --70°C in infected tissue culture fluid from the 46th tIeLa cell passage (13) (through the 1 on

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Section: Discussioncontrasting

confidence: 57%

Observations on the Fine Structure of Mature Herpes Simplex Virus and on the Composition of Its Nucleoid

Epstein

1962

The Journal of Experimental Medicine

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“…The morphology of the virus demonstrated by negative-staining techniques is consistent with its classification as an adenovirus. The apparent inner component seen in some preparations may be associated with penetration of stain and be related to the inner component described in sections of adenovirus by Epstein (1959). The general appearance and measurements of the virus in sections are consistent with fixed and embedded adenovirus (Brandon & McLean, 1962).…”

Section: Discussionsupporting

confidence: 54%

Electron microscopy of a porcine adenovirus

Chandler

Haig

Smith

1965

Journal of the Royal Microscopical Society

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SYNOPSIS A virus isolated from the pig and showing characteristics of an adenovirus has been examined by electron microscopy in both the purified state and in tissue culture cells. The virus is icosahedral in shape, has 252 capsomeres per capsid, and is characteristic of the adenovirus group. Examination of infected cells shows that the virus is predominantly intranuclear but virus has also been demonstrated in the cytoplasm. The cellular relationships characterise the adenovirus as more likely to be a member of the sub‐group containing types 3, 4 and 7 than that containing type 5. The virus is closely associated with the chromatin of the nucleus, in which parallel densely staining fibres have been demonstrated. Other structures with a rectilinear profile have been observed which have dense cores, evidence of a helical component and loci resembling the spheroidal form of virus. The first report of the isolation from a pig of a virus resembling an adenovirus was made by Haig et al. (1946): the results of serological tests indicated that the virus possessed the adenovirus group antigen but there was no neutralisation against several of the commoner human adenoviruses; the cytopathic changes in tissue culture were of the type described for the sub‐group of human adenoviruses types 3, 4 and 7. This paper records an electron‐microscope study of the morphology of the virus, the changes in infected tissue‐culture cells and the micro‐ecology of the virus.

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“…It has already been suggested that viruses differ in the fundamentals of their structure, depending on whether or not they possess a triple-layered outer limiting membrane (6), and the observations reported here indicate that this, in turn, is conditioned by the way in which a virus grows in, and leaves, the cell it parasitizes. Slow growing tumour viruses such as the Rous virus possess a triple-layered membrane (28) like that of herpes, whereas the adenovirus which bursts out of disrupted cells (29) does not (30). It might well be that the presence or absence of an outer membrane can be taken as a guide to a given agent's mode of release.…”

Section: ~Igure 10mentioning

confidence: 99%

OBSERVATIONS ON THE MODE OF RELEASE OF HERPES VIRUS FROM INFECTED HELA CELLS

Epstein

1962

The Journal of Cell Biology

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The development of a well adapted strain of herpes virus has been studied in HeLa cells using thin sectioning techniques for electron microscopy. Particular attention was directed to events in the cytoplasm and certain new features were observed. Profuse immature particles with a nucleoid and single limiting membrane were present in the nuclei of infected cells, often in crystalline array; morphologically indistinguishable immature particles were also found very frequently in the cytoplasm. Cells with such particles were intact and well preserved, and contained smooth vacuoles apparently derived from the Golgi component of the endoplasmic reticulum. The cytoplasmic particles escaped from the cells by bulging out as buds through the cell membrane or through that of the cytoplasmic vacuoles until they were attached only by a pedicle and then became free. During this process the particles were gradually enclosed by the membrane through which they passed and carried a coat of it with them as they matured. After permanganate fixation the triple-layered structure of the cell membrane and vacuolar membranes was evident and was identical with that of the outer coat of the mature virus. These findings are discussed both in relation to different types of virus structure and to function in the endoplasmic reticulum and cell membrane.Early morphological studies on herpes simplex infection using thin sectioning methods for electron microscopy (1-3) gave results which agreed on two important points. Firstly, the virus formed in the nucleus of infected cells, and appeared there as a particle having a single membrane around a dense nucleoid; and secondly, the mature extracellular virus was larger, and apparently possessed two membranes. However, the maturation process and mode of release of the agent remained obscure.The maturing particle was at first believed to acquire its second coat in the cytoplasm (1), but forms with this coat were subsequently observed within nuclei (2, 3), in groups close to the nuclear envelope, and surrounded by a membrane derived perhaps from the latter by invagination (3). More recently, elaborate infoldings and reduplications of the nuclear envelope have been found in infected cells (4, 5) and these are, at present, thought to provide a means for the virus to pass out of the intact nucleus (4), possibly by budding (5), and to be the source both of the second membrane of the agent and of the sacs known to surround it in the cytoplasm (4, 5).In the course of an investigation into the structure and composition of mature extracellular herpes virus (6), immature particles were frequently seen lying free in the cytoplasm of undamaged, morphologically intact, infected HeLa cells. This unusual finding did not seem to fit in with current views on the developmental cycle of the agent (4, 5) and it was therefore considered 589 on

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Observations on the Fine Structure of Type 5 Adenovirus

Cited by 21 publications

References 12 publications

Observations on the Fine Structure of Mature Herpes Simplex Virus and on the Composition of Its Nucleoid

Observations on the Fine Structure of Mature Herpes Simplex Virus and on the Composition of Its Nucleoid

Electron microscopy of a porcine adenovirus

OBSERVATIONS ON THE MODE OF RELEASE OF HERPES VIRUS FROM INFECTED HELA CELLS

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