Transforming Region of Group A, B, and C Adenoviruses: DNA Homology Studies with Twenty-Nine Human Adenovirus Serotypes

114

Preparation ofthe E. coli plasmid pBR322 (30). The plasmid pBR322 was prepared as described earlier (8). Extraction and purification of cellular DNA. The sodium dodecyl sulfate (SDS)-pronase B-phenol procedure was outlined in detail elsewhere (33). The DNA was also treated with RNase and proteinase K and was purified by equilibrium centrifugation in CsCl density gradients (33). Analysis of cellular DNA by the Southern blotting technique (29). The total intracellular DNA from transformed cells was cleaved with various re

Section: Giessen2 Germanymentioning

confidence: 99%

Integration Sites of Adenovirus Type 12 DNA in Transformed Hamster Cells and Hamster Tumor Cells

Stabel

Doerfler

Friis

1980

114

“…The positions and polarities of the sites complementary to early and late mRNA are similar if not identical in Ad5 and the distantly related (in terms of DNA sequence homology) Adl2 (18,23,27). This conservation of physical organization between the genomes of highly oncogenic Adl2 and nononcogenic Ad5 suggests that genes with similar functions may share common map positions.…”

mentioning

confidence: 91%

Complementation of adenovirus type 5 host range mutants by adenovirus type 12 in coinfected HeLa and BHK-21 cells

Rowe¹,

Graham²

1981

We have studied the ability of adenovirus type 12 (Adl2) to complement the Ad5 transformation-defective host range (hr) mutants during infection of human cells (HeLa) or hamster cells (BHK-21). The group I mutant hr3 (mapped within 1.3 to 3.7 map units), which is incapable of synthesizing viral DNA, was complemented for both DNA synthesis and infectious virus production in nonpermissive HeLa cells during coinfection with Adl2. Similarly, the group II mutant hr6 (6.1 to 9.4 map units), which does synthesize DNA, was also shown to be complemented for virus production. When the host cells were BHK-21, an established hamster cell line that is permissive for Ad5 but nonpermissive for Adl2 DNA synthesis and virus production, coinfection with Ad5 and Adl2 did not overcome the block to Adl2 DNA synthesis. Coinfection of BHK-21 cells with Adl2 and either hr3 or hrO leads to the complementation of only the group I mutant (hr3). The inability of Adl2 to complement hr6 in BHK-21 cells may be due to the failure of Adl2 to express an early gene product from the region corresponding to early region 1B (4.5 to 11 map units) of Ad5 where hr6 and the other group II mutations are located.

“…The human adenoviruses fall into five distinct groups based on DNA homology measurements, A through E (12a). Each group contains homologous transforming regions (22). The transforming regions of groups A (adenovirus type 12 [Adl2], Adl8, and Ad31), B (Ad3, Ad7, Adll, Adl4, Adl6, and Ad2l), and C (Adl, Ad2, Ad5, and Ad6) have been localized in early region 1 (El) in the left 1.3 to 11% of the viral genome.…”

mentioning

confidence: 99%

Identification of adenovirus 12-encoded E1A tumor antigens synthesized in infected and transformed mammalian cells and in Escherichia coli

Lucher¹,

Kimelman²,

Symington³

et al. 1984

COOH (peptide 204), targeted to the common C-terminus of human adenovirus 12 (Adl2) tumor antigens encoded by the ElA 13S mRNA and 12S mRNA, has been synthesized. Antibody prepared in rabbits against peptide 204 immunoprecipitated two proteins of apparent Mr 47,000 and 45,000 from extracts of [35S]methionine-labeled Adl2-early infected KB cells and a 47,000 protein from extracts of the Adl2-transformed hamster cell line, HE C19. Immunoprecipitation analysis of infected and transformed cells labeled with 32P, showed that both major Adl2 EIA T antigens are phosphoproteins. Immunofluorescence microscopy of Adl2-early infected KB cells with antipeptide antibody showed the site of ElA protein concentration to be predominantly nuclear. ElA proteins were detected by imtnunofluorescence at 4 to 6 h postinfection and continued to increase until at least 18 h postinfection. Antipeptide 204 antibody was used to analyze the proteins synthesized in Escherichia coli cells transformed by plasmids containing cDNA copies of the Adl2 EIA 13S mRNA or 12S mRNA under the control of the tac promoter (D.