The polypeptides of adenovirus

Maizel, Jacob V.; White, David O.; Scharff, Matthew D.

doi:10.1016/0042-6822(68)90121-9

Cited by 763 publications

(134 citation statements)

References 20 publications

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“…The sources of HeLa cells, Ad2, and Ad5 have been described (23,24 [3H]dTTP, the other three deoxynucleotides, and ATP that had been warmed to the temperature indicated for the reaction. All radioactive labeling was measured after 120 min.…”

Section: Methodsmentioning

confidence: 99%

“…Channel A contains the supernate from Ad2-infected cells from which the Ad2 DBP was purified. Roman numerals refer to the nomenclature of virion proteins proposed by Maizel et al (23). The purity of the DBP polypeptide was measured by densitometry of the Coomasie blue-stained wet gel on a Transidyne General Corporation scanning densitometer equipped with a digital computing integrator.…”

Section: Methodsmentioning

confidence: 99%

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Temperature-sensitive replication of H5ts125 adenovirus DNA in vitro.

Horwitz¹

1978

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

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A soluble adenovirus DNA replication system has been prepared in extracts of HeLa cells infected with the temrperature-sensitive mutant HMts125. These nuclear extracts synthesize full-sized viral DNA at 300 but are temperature sensitive in this function at 380. A 72,000-dalton DNA-binding protein purified from cells infected with wild-type virus complements the temperature-sensitive defect and restores viral DNA replication to 74% of normal values. We have recently described the preparation of a nuclear extract from adenovirus (Ad)infected cells that catalyzed the synthesis of viral DNA in a manner analogous to that used in whole cells (1). These extracts, which were free of chromosomal DNA, contained replicating Ad DNA and synthesized full-size viral DNA products. Similar approaches developed in prokaryotes and their infecting bacteriophages have allowed the analysis of various gene products required for DNA initiation and elongation (2)(3)(4)(5). The general approach has been to prepare extracts from bacterial cell mutants that are temperature sensitive (ts) in DNA synthesis and to use these extracts as a receptor to isolate biochemically complementing activity from wild-type

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Temperature-sensitive replication of H5ts125 adenovirus DNA in vitro.

Horwitz¹

1978

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

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“…Stocks of adenovirus type 2 were prepared and stored as described by Maizel et al (6). Virus preparations were used within 1 month and were free of identifiable, small, adeno-associated virus.…”

Section: Methodsmentioning

confidence: 99%

Adenovirus Messenger RNA in Mammalian Cells: Failure of Polyribosome Association in the Absence of Nuclear Cleavage

McGuire

Swart

Hodge

1972

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

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The nuclear synthesis of adenovirus-specific RNA late in the infectious cyclein the presence of toyocamycin (an adenosine analogue) has been investigated. There is reduced synthesis of viral RNA with an accumulation of virus-specific RNA in the molecular weight range of at least 4 to 8 X 106. No new viral RNA associates with cytoplasmic polyribosomes. In addition, hybridization competition experiments indicate a 70% competition between these large nuclear transcripts and polyribosomeassociated viral RNA that was synthesized in the absence of inhibitor. These data are consistent with the following interpretations: complete nuclear processing of viral RNA is necessary for polyribosome association, and precursor viral message(s) contain sequences that are lost normally during post-transcriptional processing.Evidence has been presented that messneger RNA(mRNA) in mammalian cells may undergo a process of cleavage after transcription. A precursor-product relationship has been suggested between nuclear heterogeneous RNA of high molecular weight and the mRNA of lower molecular weight that is associated with polysomes in HeLa cells (1). In addition, cells either lytically infected or transformed by DNA nuclear viruses contain larger virus-specific RNA transcripts in the nucleus than in the cytoplasm (2, 3). To examine in more detail the nature of this cleavage of mRNA and its possible implications in mammalian cells, it would be helpful if the processes of synthesis and cleavage could be separated. This separation has been achieved by the use of toyocamycin in studies on processing of ribosomal RNA (4). When used in relatively low concentrations, this inhibitor is incorporated into the newly synthesized 45S RNA resulting in the subsequent failure of processing. In the present study, the effect of toyocamycin on processing of viral RNA has been investigated. HeLa cells, at 22 hr after infection with adenovirus 2, were incubated with this inhibitor and the synthesis, accumulation, and fate of nuclear viral transcripts were ascertained. The data demonstrate that in the presence of inhibitor large virus-specific transcripts are synthesized and accumulate in the nucleus, but fail to be processed. Under these conditions, no virus-specific RNA becomes associated with cytoplasmic polyribosomes. MATERIALS AND METHODSMaintenance of Cells. HeLa S3 cells were maintained in suspension culture in Eagle's medium (5) supplemented with 7% calf serum.Preparation of Virus and Infection of Cells. Stocks of adenovirus type 2 were prepared and stored as described by Maizel et al. (6). Virus preparations were used within 1 month and were free of identifiable, small, adeno-associated virus. Cells were infected at a concentration of 2000 viral particles per cell as described (7).Preparation of Nuclear RNA. Nucleoplasm was prepared from HeLa cells by the procedure of Penman (8). Nucleic acid and protein were precipitated at -20°for 16 hr with 0.15 M sodium chloride, 0.1% of 2-mercaptoethanol, and 2.5 volumes of ethanol. This precipitate ...

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“…OBP-301 and OBP-405 viruses were purified by CsCl 2 step gradient ultracentrifugation followed by CsCl 2 linear gradient ultracentrifugation. Determination of virus particle titer and infectious titer was accomplished spectrophotometrically by the method of Maizel et al (1968) and by the method of Kanegae et al (1994), respectively.…”

Section: Recombinant Adenovirusesmentioning

confidence: 99%

Enhanced oncolysis by a tropism-modified telomerase-specific replication-selective adenoviral agent OBP-405 (‘Telomelysin-RGD’)

et al. 2005

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Replication-competent oncolytic viruses are being developed for human cancer therapy. We previously reported that an attenuated adenovirus (OBP-301, 'Telomelysin'), in which the hTERT promoter element drives expression of E1A and E1B genes linked with an IRES, could replicate in cancer cells, and causes selective lysis of cancer cells. We further constructed OBP-405 ('Telomelysin-RGD') that contains an RGD motif in the HI loop of the fiber knob. We examined whether OBP-405 could be effective in overcoming the limitations of OBP-301, specifically their inefficient infection into cells lacking the primary receptor, the coxsackievirus and adenovirus receptor (CAR). By flow cytometric analysis, H1299 (lung) and SW620 (colorectal) tumor cells showed high levels of CAR expression, whereas LN444 (glioblastoma), LNZ308 (glioblastoma), and H1299-R5 (lung) tumor cells were negative for CAR expression. A quantitative realtime PCR analysis demonstrated that fiber-modified OBP-405 infected more efficiently than OBP-301, although the intracellular replication rate of both viruses was consistent. The comparative antitumor effect of fibermodified OBP-405 and unmodified OBP-301 for human cancer cells was evaluated in vitro by XTT assay as well as in vivo by using athymic mice carrying xenografts. OBP-405 had a profound oncolytic effect on human cancer cell lines compared to OBP-301, in particular on cells with low CAR expression. Intratumoral injection of 10 7 plaque-forming units of OBP-405 into CAR-negative H1299-R5 lung tumor xenografts in nu/nu mice resulted in a significant inhibition of tumor growth and long-term survival in all treated mice. Moreover, selective replication of OBP-405 in the distant, uninjected H1299-R5 tumors was demonstrated. Our results suggest that fiber-modified replication-competent adenovirus OBP-405 exhibits a broad target range by increasing infection efficiency, an outcome that has important implications for the treatment of human cancers.

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The polypeptides of adenovirus

Cited by 763 publications

References 20 publications

Temperature-sensitive replication of H5ts125 adenovirus DNA in vitro.

Temperature-sensitive replication of H5ts125 adenovirus DNA in vitro.

Adenovirus Messenger RNA in Mammalian Cells: Failure of Polyribosome Association in the Absence of Nuclear Cleavage

Enhanced oncolysis by a tropism-modified telomerase-specific replication-selective adenoviral agent OBP-405 (‘Telomelysin-RGD’)

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