Mapping of base sequence heterologies between genomes from different adenovirus serotypes

Garon, Claude F.; Berry, Karen W.; Hierholzer, John C.; Rose, James A.

doi:10.1016/0042-6822(73)90153-0

Cited by 95 publications

(26 citation statements)

References 4 publications

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“…To demonstrate that the template for in vitro synthesis is the exogenously added DNA, we also analyzed the product synthesized by AdS-infected-cell extract in response to DNA-protein complex prepared from adenovirus 2 (Ad2) virions. The extensive homology between the genomes of Ad2 and AdS (28) suggested that the two DNAs should be interchangeable as templates for the in vitro reaction, and such was in fact the case. The rate of DNA synthesis in AdS-infected-cell extract was virtually identical with either Ad5 or Ad2 DNA-protein complex as substrate (data not shown).…”

Section: Methodsmentioning

confidence: 99%

Adenovirus DNA replication in vitro.

Challberg¹,

Kelly²

1979

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

244

119

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Much of the recent dramatic progress in understanding prokaryotic DNA replication is due to the development of soluble in vitro systems that carry out the replication of exogenously added DNA templates of defined structure (1-4). These systems have made it possible to study in detail the biochemical mechanisms involved in the initiation and elongation of DNA chains and to purify and characterize the proteins required for these processes. It is clear that the development of analogous in vitro systems would be extremely valuable for dissecting the mechanisms of eukaryotic DNA replication.The human adenoviruses have several advantages as model systems for studying eukaryotic DNA replication. The adenovirus genome is easily isolated in intact form from purified virions, and its structure and genetic organization are well defined (5, 6). In addition, a good deal is known about the general mechanism of adenovirus DNA replication in vivo (7). This makes it possible to assess the biological relevance of any synthetic reactions observed in vitro. The adenovirus genome is a linear, duplex DNA molecule with a molecular weight of 20-25 X 106 (8). It possesses two novel structural features of possible significance for DNA replication. First, the two ends of the genome have identical nucleotide sequences (refs. 9-11; J. R. Arrand and R. J. Roberts, personal communication), and second, the 5' ends of both DNA strands are covalently linked to a polypeptide of molecular weight 55,000 (12)(13)(14)(15)(16). In vivo studies (reviewed in ref. 7) have shown that initiation of adenovirus DNA replication takes place at or near either end of the viral genome. Daughter strand synthesis then proceeds in the 5' to 3' direction with concomitant displacement of the parental strand with the same polarity. Synthesis of the complementary strand is initiated at or near the 3' end of the displaced parental strand and also proceeds in the 5' to 3' direction. Thus, the overall pattern of adenovirus DNA replication appears ratherThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance' with 18 U. S. C. §1734 solely to indicate this fact. 655 simple. At each growing point only one of the two parental strands is replicated, and, at least in principle, there is no requirement for a discontinuous mechanism of DNA synthesis. This simplicity makes the adenovirus system especially attractive for biochemical studies.In this paper we describe a soluble enzyme system from adenovirus-infected cells that is capable of replicating exogenously added adenovirus DNA molecules. We have shown by several criteria that the DNA synthesis that occurs in this in vitro system closely resembles adenovirus DNA replication in vivo. METHODSPreparation of Nuclei from Adenovirus-Infected Cells. HeLa cells were maintained at 370C in suspension culture in Eagle's minimal essential medium supplemented with horse serum (5%). One liter of cells at a density of 4-5 X 105 cells/ml was ...

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Section: Methodsmentioning

confidence: 99%

Adenovirus DNA replication in vitro.

Challberg¹,

Kelly²

1979

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

244

119

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“…By heteroduplex mapping of genomes from different viral serotypes, early region 3, in which the E19 protein is encoded (8)(9)(10), has been shown to be one of two regions of the adenovirus genome that exhibits significant divergence even within subgenera (11,12). Furthermore, early region 3 has been shown to be dispensable for the virus when grown in vitro (13).…”

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confidence: 99%

Adenoviruses of subgenera B, C, D, and E modulate cell-surface expression of major histocompatibility complex class I antigens.

Pääbo¹,

Nilsson²,

Peterson³

1986

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

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“…The region responsible for the transforming capacity has been demonstrated to be restricted to only 6% at the left end of the adenovirus type 5 (Ad 5) genome [3]. In spite of this, the division of adenoviruses into subgroups A to D has been useful, since additional properties are shared between subgroup members: the GC content of the viral DNA [4]; the sequence homology detected by filter hybridization [4] or heteroduplex mapping [5]: differences in the length of the vertex projections of adenovirions [6]: the form of the soluble hemagglu tinin [6,7]; antigenic relationships of struc tural and nonstructural antigens [8][9][10][11]: and also differences in the pathogenicity of differ ent serotypes [12,13].…”

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confidence: 99%

Classification of Human Adenoviruses by SDS-Polyacrylamide Gel Electrophoresis of Structural Polypeptides

Wadell¹

1979

Intervirology

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The virion polypeptide compositions of 15 human adenovirus types, representing all members of subgroups B and C and selected members of subgroups A and D, have been analyzed by SDS-PAGE. The capsid polypeptides II, III, Ilia and IV displayed a pattern of apparent molecular weights which was characteristic for each adenovirus serotype analyzed. Adenovirus serotypes belonging to the same subgroup were also demonstrated to share an apparent molecular weight pattern of the internal structural polypeptides in the following manner: Subgroup A: V – resolved into two bands of 46–48.5 K and 51–51.5 K; VI – 25.5 K; and VII- 18 K. Subgroup B: V- 53.5–54.5 K; VI-24 K; and VII – 18 K. Subgroup C: V – 48.5 K; VI – 24 K; and VII – 18.5 K. Subgroup D: V – 50.5 K; VI – 23.5 K; and VII – 18 K. Adenovirus type 4: V – 48 K; VI – 24.5 K; and VII – 18 K. The structural polypeptides represent a major portion of the adenovirus gene products. It is therefore proposed that the apparent molecular weight differences of viral polypeptides be used (i) to confirm the identification of an adenovirus type, and (ii) for subgroup classification of adenoviruses and, consequently, that adenovirus type 4 should be classified as a member of a separate subgroup designated E.

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Mapping of base sequence heterologies between genomes from different adenovirus serotypes

Cited by 95 publications

References 4 publications

Adenovirus DNA replication in vitro.

Adenovirus DNA replication in vitro.

Adenoviruses of subgenera B, C, D, and E modulate cell-surface expression of major histocompatibility complex class I antigens.

Classification of Human Adenoviruses by SDS-Polyacrylamide Gel Electrophoresis of Structural Polypeptides

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