Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
DNA methylation of three 5' C-C-G-G 3' sites in the promoter and 5' region inactivate the E2a gene of adenovirus type 2.
Langner KD
,
Vardimon L
,
Renz D
,
Doerfler W
.
???displayArticle.abstract???
The E2a gene of human adenovirus type 2 (Ad2) encodes the 72-kilodalton DNA-binding protein. We previously described perfect inverse correlations between the methylation of all 5' C-C-G-G 3' sequences in the Ad2 E2a gene in virus-transformed hamster cells containing viral DNA sequences in an integrated state and the extent to which this gene is expressed. We subsequently showed that in vitro methylation of all 14 5' C-C-G-G 3' sequences in the cloned E2a gene by prokaryotic Hpa II DNA methyltransferase leads to transcriptional inactivation after microinjection into Xenopus laevis oocytes. The unmethylated cloned E2a gene is expressed in these cells. We report here the construction of partly methylated clones of the E2a gene. In the promoter (5')-methylated construct, three 5' C-C-G-G 3' sequences at the 5' end of the subclone were methylated. One of these sites is located 215 base pairs (bp) upstream (bp 26,169 of Ad2 DNA), and two sites are located 5 and 23 bp downstream from the cap site (bp 25,931 and 25,949 of Ad2 DNA) of the E2a gene. This construct was transcriptionally inactive upon microinjection into nuclei of X. laevis oocytes. In the gene (3')-methylated construct, 11 5' C-C-G-G 3' sequences in the main part of the transcribed gene region were methylated in vitro. This construct was transcribed in X. laevis oocytes, and at least some of the Ad2-specific RNA synthesized was initiated at the same sites as in Ad2-infected human KB cells. Both mock-methylated constructs were transcribed into Ad2-specific RNA in X. laevis oocytes. These results demonstrate that DNA methylations at or close to the promoter and 5' end of the E2a gene cause transcriptional inactivation. Perhaps only one methyl group would be adequate for inactivation; in vivo methylation of more than one cytosine may be a form of safeguard or redundancy.
Alwine,
Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes.
1977, Pubmed
Alwine,
Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes.
1977,
Pubmed
Baker,
Promoters and heterogeneous 5' termini of the messenger RNAs of adenovirus serotype 2.
1981,
Pubmed
BARTH,
Differentiation of cells of the Rana pipiens gastrula in unconditioned medium.
1959,
Pubmed
Behe,
Effects of methylation on a synthetic polynucleotide: the B--Z transition in poly(dG-m5dC).poly(dG-m5dC).
1981,
Pubmed
Berk,
Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids.
1977,
Pubmed
Busslinger,
DNA methylation and the regulation of globin gene expression.
1983,
Pubmed
Constantinides,
Functional striated muscle cells from non-myoblast precursors following 5-azacytidine treatment.
1977,
Pubmed
Doerfler,
DNA methylation and its functional significance: studies on the adenovirus system.
1984,
Pubmed
Doerfler,
DNA methylation--a regulatory signal in eukaryotic gene expression.
1981,
Pubmed
Doerfler,
DNA methylation and gene activity.
1983,
Pubmed
Ehrlich,
5-Methylcytosine in eukaryotic DNA.
1981,
Pubmed
Flavell,
Structure and expression of the human globin genes and murine histocompatibility antigen genes.
1983,
Pubmed
Fradin,
Methylation of simian virus 40 Hpa II site affects late, but not early, viral gene expression.
1982,
Pubmed
,
Xenbase
Gorman,
Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells.
1982,
Pubmed
Graham,
A new technique for the assay of infectivity of human adenovirus 5 DNA.
1973,
Pubmed
Groudine,
Chromatin structure of endogenous retroviral genes and activation by an inhibitor of DNA methylation.
1981,
Pubmed
Haigh,
DNA methylation in chicken alpha-globin gene expression.
1982,
Pubmed
Kruczek,
The unmethylated state of the promoter/leader and 5'-regions of integrated adenovirus genes correlates with gene expression.
1982,
Pubmed
,
Xenbase
Kruczek,
Expression of the chloramphenicol acetyltransferase gene in mammalian cells under the control of adenovirus type 12 promoters: effect of promoter methylation on gene expression.
1983,
Pubmed
La Volpe,
Coupled demethylation of sites in a conserved sequence of Xenopus ribosomal DNA.
1983,
Pubmed
,
Xenbase
Ley,
5-azacytidine selectively increases gamma-globin synthesis in a patient with beta+ thalassemia.
1982,
Pubmed
Macleod,
Transcription in oocytes of highly methylated rDNA from Xenopus laevis sperm.
,
Pubmed
,
Xenbase
Mann,
Specificity of Hpa II and Hae III DNA methylases.
1977,
Pubmed
Mohandas,
Reactivation of an inactive human X chromosome: evidence for X inactivation by DNA methylation.
1981,
Pubmed
Ott,
Undermethylation at the 5' end of the albumin gene is necessary but not sufficient for albumin production by rat hepatoma cells in culture.
1982,
Pubmed
Radloff,
A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells.
1967,
Pubmed
Razin,
DNA methylation and gene function.
1980,
Pubmed
Rigby,
Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I.
1977,
Pubmed
Riggs,
5-methylcytosine, gene regulation, and cancer.
1983,
Pubmed
Schirm,
Expression of viral DNA in adenovirus type 12-transformed cells, in tumor cells, and in revertants.
1981,
Pubmed
Simon,
Retrovirus genomes methylated by mammalian but not bacterial methylase are non-infectious.
,
Pubmed
Southern,
Detection of specific sequences among DNA fragments separated by gel electrophoresis.
1975,
Pubmed
Stabel,
Integration sites of adenovirus type 12 DNA in transformed hamster cells and hamster tumor cells.
1980,
Pubmed
Stein,
In vitro methylation of the hamster adenine phosphoribosyltransferase gene inhibits its expression in mouse L cells.
1982,
Pubmed
Stein,
Pattern of methylation of two genes coding for housekeeping functions.
1983,
Pubmed
Sutter,
Methylation of integrated viral DNA sequences in hamster cells transformed by adenovirus 12.
1980,
Pubmed
Sutter,
Methylation of integrated adenovirus type 12 DNA sequences in transformed cells is inversely correlated with viral gene expression.
1980,
Pubmed
Taylor,
Multiple new phenotypes induced in 10T1/2 and 3T3 cells treated with 5-azacytidine.
1979,
Pubmed
Vardimon,
In vitro methylation of the BsuRI (5'-GGCC-3') sites in the E2a region of adenovirus type 2 DNA does not affect expression in Xenopus laevis oocytes.
1982,
Pubmed
,
Xenbase
Vardimon,
DNA methylation and viral gene expression in adenovirus-transformed and -infected cells.
1980,
Pubmed
Vardimon,
Expression of a cloned adenovirus gene is inhibited by in vitro methylation.
1982,
Pubmed
,
Xenbase
Vardimon,
Methylation of adenovirus genes in transformed cells and in vitro: influence on the regulation of gene expression?
1981,
Pubmed
,
Xenbase
Waalwijk,
MspI, an isoschizomer of hpaII which cleaves both unmethylated and methylated hpaII sites.
1978,
Pubmed
Waechter,
Effect of methylation on expression of microinjected genes.
1982,
Pubmed
Wahl,
Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate.
1979,
Pubmed
Wilks,
Estrogen induces a demethylation at the 5' end region of the chicken vitellogenin gene.
1982,
Pubmed
