Controlled degradation of vaccinia virions in vitro: an electron microscopic study

Easterbrook, K. B.

doi:10.1016/s0022-5320(66)80077-1

Cited by 136 publications

(82 citation statements)

References 16 publications

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“…The preparation was according to the procedure of Easterbrook (1966). Purified modified vaccinia virus Ankara (MVA, 2000 ~tg) was resnspended in phosphatebuffered saline (PBS), dialysed and incubated with 1 ~ NP40 and 50 Ixl 2-mercaptoethanol for 60 min at 37 °C.…”

Section: Methodsmentioning

confidence: 99%

Structural and functional analysis of orthopoxvirus epitopes with neutralizing monoclonal antibodies

Czerny¹,

Mahnel²

1990

Journal of General Virology

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Section: Methodsmentioning

confidence: 99%

Structural and functional analysis of orthopoxvirus epitopes with neutralizing monoclonal antibodies

Czerny¹,

Mahnel²

1990

Journal of General Virology

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“…Evidence has been presented that the isolated material is specifically associated with DNA. Structural studies of vaccinia virus particles indicate that the DNA of the virus genome is contained within enclosed cores under the outer coat of the virus particle (Easterbrook, 1966). Studies using radioactively labelled rabbit pox virus have shown that, although the virus genome is not digested by exposure of the virus to deoxyribonuclease, small amounts of DNA are released from the surface of the virus particle.…”

Section: Isolation Of a Vaccinia-specific Hapten 367 Discussionmentioning

confidence: 99%

Isolation of a Vaccinia-specific Hapten from Vaccinia-infected Sheep Dermis

Williamson¹,

Rondle

1968

Journal of General Virology

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SUMMARYVirus-free buffer extracts of vaccinia-infected sheep dermis were fractionated by elution from DEAE-cellulose with increasing concentrations of NaC1. The fraction eluted with 4-o M-NaC1 was serologically homogeneous when examined in precipitation-in-gel tests. Further fractionation on Sephadex G-2oo gave a material which was also physically homogeneous when examined by both ultracentrifugal and electrophoretic techniques. A sedimentation coefficient of 3"47 S and a molecular weight of 3 I,OOO were calculated from ultracentrifugal data. The isolated material contained 74"3 % DNA, 9"4 % protein estimated as bovine serum albumin and 14"9 % carbohydrate estimated as glucose. The DNA contained the bases adenine, thymine, guanine and cytosine in the ratios I.O: I. t : o.8: o.8. Paper chromatography of formic-acid hydrolysates of the material resolved seven substances reacting with ninhydrin. Qualitative colorimetric tests indicated, apart from deoxyribose, the presence of a hexose and a hexuronic acid.The serological activity of the isolated material was heat-stable and identical with that of a previously described heat-stable extract of vacciniainfected rabbit dermis. Treatment with enzymes indicated the presence of two different, serologically active sites. Failure to elicit an antibody response following injection into rabbits suggested that the isolated material was a hapten. Serum absorption studies showed serological identity with an antigen present on the surface of the vaccinia virus particle.

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“…The level of this inhibition depended on the concentration of the vaccinia cores. A 50% decrease of protein synthesis (relative to the control) was obtained with 10 ,l of cores, corresponding to 300 Mug of viral protein per ml (Fig. 2).…”

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confidence: 82%

“…175 crose density gradient centrifugation as described (9) except that an additional cycle of sucrose cushion followed by sucrose gradient centrifugation was performed. Cores were prepared by incubation of purified virus (previously sonicated for 40 sec at 4 ,u on an MSE sonicator) at 370 for 30 min in 30 mM TrisHCl, pH 7.5/0.2% Triton X-100 (Koch-Light)/50 mM 2-mercaptoethanol (10). At the end of this incubation, the Triton X-100 was removed from the sonicated cores by adsorption on Bio-Beads SM2 (Bio-Rad) (11) once in ethanol for 5 min, in ethanol/diethyl ether, 1:1 (vol/vol), and finally in diethyl ether.…”

mentioning

confidence: 99%

“…The wheat germ cell-free extract was prepared essentially according to the procedure of Davies and Kaesberg (13). The standard in vitro reaction mixture for amino acid incorporation directed by TYMV RNA (80 ,g/ml) consisted of 100 ,ul of a reaction mixture containing 20 mM N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (Hepes) (pH 7.6), 3 Mg/ml, cycloheximide at 300 ,g/ml, and [14C]phenylalanine (10 ,Ci/ml, 370 mCi/mmol). Incubation was at 300 for 50 min.…”

mentioning

confidence: 99%

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In vitro inhibition of protein synthesis by purified cores from vaccinia virus.

Ben-Hamida¹,

Beaud²

1978

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

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The mechanism of the shutoff of cellular protein synthesis in vaccinia virus-infected cells has been investigated by using in vitro systems. Purified vaccinia cores cause inhibition of endogenous mRNA translation in nonpreincubated reticulocyte lysates and Ehrlich ascites tumor cell-free systems. Translation of viral mRNA from turnip yellow mosaic virus is also impaired in wheat germ cell-free extracts. The block induced by vaccinia cores in protein synthesis is not due to a decrease in the availability of mRNA but rather to an alteration of the cellular translational machinery. No nucleolytic activity able of digesting mRNA could be detected in purified vaccinia cores with three sensitive tests. There is a lack of inhibition in the poly(Phe)poly(U) system, which bypasses the normal initiation process. An almost complete disaggregation of polyribosomes in the reticulocyte lysate appears when vaccinia cores are present. These results indicate that mRNA translation in a cell-free system is affected predominantly at the level of polypeptide chain initiation.

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Controlled degradation of vaccinia virions in vitro: an electron microscopic study

Cited by 136 publications

References 16 publications

Structural and functional analysis of orthopoxvirus epitopes with neutralizing monoclonal antibodies

Structural and functional analysis of orthopoxvirus epitopes with neutralizing monoclonal antibodies

Isolation of a Vaccinia-specific Hapten from Vaccinia-infected Sheep Dermis

In vitro inhibition of protein synthesis by purified cores from vaccinia virus.

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