Adenovirus type 2 DNA replication. II. Termini of DNA replication

Schilling, R; Weingärtner, B; Winnacker, Ernst‐Ludwig

doi:10.1128/jvi.16.4.767-774.1975

Cited by 46 publications

(13 citation statements)

References 19 publications

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“…This labeling pattern clearly supports bidirectional growth of Ad2 DNA when labeled in vitro in the intact soluble replication complex. These patterns very closely resemble those of others which were generated by a similar analysis of completed DNA pulse-labeled for short times in vivo (2,5,11,15,16,17).…”

supporting

confidence: 83%

“…The numbers beneath fragment designations refer to the position of the fragment within the Ad2 genome. now well established (2,5,11,15,16,17). In the current work we have extended the general method of Danna and Nathans (4) to the in vitro observation of replication termini, thus further strongly supporting the contention of in vitro termination within the Ad2 soluble replication system.…”

mentioning

confidence: 99%

“…The design of the experiments described herein is basically that of Danna and Nathans (4), as used to demonstrate bidirectional replication of SV40 DNA. This general technique has been applied to demonstrate termination sites in Ad2 DNA labeled in vivo (2,5,11,15,16) and has now been further extended to the analysis of Ad2 DNA labeled in vitro by the soluble replication complex. Since the types of experiments undertaken to demonstrate in vitro termination require that complete 31S doublestranded (ds) DNA be made, it was necessary to show that each soluble complex used synthesized full-length Ad2 DNA during the in vitro reaction.…”

mentioning

confidence: 99%

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In vitro termination of adenovirus DNA synthesis by a soluble replication complex

Arens¹,

Yamashita²

1978

J Virol

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The double-stranded DNA from a soluble DNA replication complex that was labeled with deoxyribonucleoside triphosphates and completed in vitro was digested with EcoRI, Sma I, and Hpa I restriction endonucleases. All regions of the adenovirus type 2 genome were labeled in vitro, but restriction fragments derived from the ends of the DNA molecules were relatively more highly labeled than those derived from internal regions. The in vitro endogenous DNA polymerase reaction also exhibited strand-specific labeling near the molecular ends, in that restriction fragments from the left end were labeled predominantly in the r strand and fragments from the right end were labeled predominantly in the I strand.

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supporting

confidence: 83%

mentioning

confidence: 99%

mentioning

confidence: 99%

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In vitro termination of adenovirus DNA synthesis by a soluble replication complex

Arens¹,

Yamashita²

1978

J Virol

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“…Pulse-chase labeling of DNA. The "temperatureshift" procedure described by Schilling et al (20) was applied. KB cells productively infected with 2,000 Ad2 particles/cell were sedimented at 40C and resuspended in cold Eagle MEM at a cell density of 107 cells/ml.…”

mentioning

confidence: 99%

“…Equilibrium density centrifugation. Preparative gradients of Metrizamide (Nyegaard and Co., A/S, Oslo, Norway) were made by mixing 6.5 ml each of 20 and 60% (wt/vol) Metrizamide in 0.01 M Trishydrochloride buffer (pH 8.1), 1 mM EDTA, and 0.5% (vol/vol) Triton X-100 (20 to 60% MAT gradients). Metrizamide solutions were freshly made, and the gradients were stored overnight at 4C, shielded from light, and used the next day.…”

mentioning

confidence: 99%

Synthesis and processing of the precursor to the major core protein of adenovirus type 2

Everitt¹,

Meador²,

Levine³

1977

J Virol

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An isopycnic Metrizamide-detergent gradient system was developed in which the newly synthesized precursor (polypeptide P-VII) to the major core protein of adenovirus type 2 (polypeptide VII) was confined to a spectrum of complexes with densities equal to or higher than that of adenovirions. The majority of the newly synthesized P-VII was, at the beginning of the logarithmic period of virus production, present as an entity of protein density. This pool of P-VII was efficiently depleted. P-VII was also associated with high-molecular-weight structures of intermediate density, sharing some properties with empty capsids or incomplete particles. The transfer of P-VII from the intermediate-density region was not quantitative, and only particles of true virion density subsequently contained polypeptide VII. No structures equivalent to the core structure of disrupted virions or identical to incomplete particles were detected in this system. A temperature-dependent transition of radioactivity from polypeptide P-VII into polypeptide VII was also detectable after in vitro incubation of P-VIIcontaining complexes. Addition of Ad2-infected cell extracts was required for processing of complexes derived from regions of protein density, whereas P-VII was processed spontaneously upon incubation in complexes of virion density.

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Defective expression of mouse adenovirus Fl in human cells.

Antoine¹,

Aleström²,

Schilling³

et al. 1982

The EMBO Journal

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HeLa cells infected with mouse adenovirus strain Fl (AdFl) produce at least 2000 times less virus than permissive mouse 3T3 cells. Viral DNA synthesis, however, proceeds unimpaired. The defect in virion production was linked to a dramatic reduction in the synthesis of AdFI structural proteins, in particular the hexon. The identity of the AdFl hexon gene product was recognized through its immunogenic reactivity towards an antiserum raised against the human adenovirus type 2 (Ad2) hexon gene product. This crossreactivity is reflected in an extensive DNA sequence homology between AdFI and Ad2 DNA at position 53-60, the locus of the hexon gene, on the Ad2 physical map. Through hybridization at different formamide concentrations, the present study identifies one additional, highly conserved region at map positions 14-15 on the Ad2 genome.

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Adenovirus type 2 DNA replication. II. Termini of DNA replication

Cited by 46 publications

References 19 publications

In vitro termination of adenovirus DNA synthesis by a soluble replication complex

In vitro termination of adenovirus DNA synthesis by a soluble replication complex

Synthesis and processing of the precursor to the major core protein of adenovirus type 2

Defective expression of mouse adenovirus Fl in human cells.

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