+ Translate

Expression of transferred thymidine kinase genes is controlled by methylation

, : Expression of transferred thymidine kinase genes is controlled by methylation. Proceedings of the National Academy of Sciences of the United States of America 79(20): 6299-6303

Plasmid pTKx-1, containing the herpes simplex virus gene for thymidine kinase (TK) inserted into the BamHI site of plasmid pBR322, was introduced into LtK- [thymidine kinase deficient mouse fibroblasts] cells by calcium phosphate precipitation in the absence of carrier DNA. Line 101 is a TK+ derivative of Ltk- that contains multiple copies of pTKx-1 in a multimeric structure. A derivative of 101 that retained but no longer expressed the herpes simplex TK genes (termed 101BU1) and derivatives of line 101BU1 that reexpressed the genes (termed 101H1, 101HC and 101HG) were selected. The TK genes in 101BU1 were hypermethylated relative to those in the TK+ parent and derivatives. Growth of 101BU1 in the presence of the methylation inhibitor 5-azacytidine resulted in an average 13-fold increase in the number of TK+ reexpressors. DNA from 101BU1 was inactive in secondary gene transfer, whereas DNA from 101 and from TK+ reexpressors was active. These data support a causative relationship between DNA methylation and decreased gene expression. All TK+ reexpressors examined had DNA rearrangements involving TK DNA.

Accession: 005432329

PMID: 6183659

DOI: 10.2307/12861

Download PDF Full Text: Expression of transferred thymidine kinase genes is controlled by methylation

Submit PDF Full Text

No spam - Every submission is manually reviewed

Due to poor quality, we do not accept files from Researchgate

Submitted PDF Full Texts will always be free for everyone
(We only charge for PDFs that we need to acquire)

Select a PDF file:

Related references

Moolten, F.L.; Wells, J.M., 1990: Curability of tumors bearing herpes thymidine kinase genes transferred by retroviral vectors. Retroviral vectors constructed to contain the herpes simplex virus thymidine kinase (HSV-TK) gene were used for transduction of this gene into murine sarcoma and lymphoma cells to yield sublines susceptible in vitro to the cytotoxicity of ganciclo...

Christy, B.A.; Scangos, G.A., 1984: Changes in structure and methylation pattern in a cluster of thymidine kinase genes. Cell line 101 is a thymidine kinase (TK)-positive derivative of Ltk- which contains ca. 20 copies of the herpes simplex virus TK gene organized in a tandem array. DNA methylation at three sites within the gene and flanking sequences was inversely...

Christy B.; Scangos G., 1984: Dna methylation controls expression of transferred genes in a mouse l cell derivative. Pearson, M L And N L Sternberg (Ed ) Progress in Cancer Research And Therapy, Vol 30 Gene Transfer And Cancer; 4th Litton Technology Transfer Workshop, Apr 1982 Xxi+376p Raven Press: New York, N Y , Usa Illus P259-264

Scott, S.D.; Ross, N.L.; Binns, M.M., 1989: Nucleotide and predicted amino acid sequences of the Marek's disease virus and turkey herpesvirus thymidine kinase genes; comparison with thymidine kinase genes of other herpesviruses. In this paper we present the nucleotide sequences of the thymidine kinase (TK) genes of two avian herpesviruses: a highly oncogenic strain of Marek's disease virus (MDV strain RBIB) and its serologically related vaccine virus, the herpesvirus...

Hardies S.C.; Axelrod D.E.; Edgell M.H.; Hutchison C.A.IIi, 1984: Concerted hyper methylation and stable shutdown of a cluster of thymidine kinase genes. Pearson, M L And N L Sternberg (Ed ) Progress in Cancer Research And Therapy, Vol 30 Gene Transfer And Cancer; 4th Litton Technology Transfer Workshop, Apr 1982 Xxi+376p Raven Press: New York, N Y , Usa Illus P249-258

Dobrovic, A.; Gareau, J.L.; Ouellette, G.; Bradley, W.E., 1988: DNA methylation and genetic inactivation at thymidine kinase locus: two different mechanisms for silencing autosomal genes. Patterns of methylation of CpG dinucleotides in the promoter region of the thymidine kinase (TK) gene in wild-type and TK-deficient Chinese hamster cell lines were studied. Whereas wild-type cells were unmethylated, three conventionally derived TK...

Tasseron J.G.; Den Dulk H.; Van D.P.tte P.; Giphart Gassler M., 1989: De novo methylation as major event in the inactivation of transfected herpesvirus thymidine kinase genes in human cells. Spontaneous inactivation of integrated thymidine kinase genes was studied in three human cell lines, one with multiple copies and two with a single copy of a transfected shuttle plasmid containing two selectable genes: the HSV tk gene and the Eco...

Marx, J.L., 1983: Specific expression of transferred genes. Foreign genes, which were transferred into mice, appear to be expressed according to more normal patterns of tissue distribution. Science 222(4627): 1001-1002

DeVries, P.J.; Davidson, R.L.; Clough, D.W., 1986: Site specificity of DNA methylation and expression of herpes simplex thymidine kinase gene. Analysis of the methylation pattern of a single-copy herpes simplex virus thymidine kinase gene integrated into the genome of mouse L cells revealed that hypomethylation of five specific AvaI sites correlates with expression of the TK gene in all...

Glebov, O.K.; Abramyan, D.S., 1985: Genetic transformation of somatic cells viii. effect of dna methylation inhibitor 5 azacytidine on the stability of thymidine kinase negative and thymidine kinase positive phenotype of transformant clone cells. A study was made of the effect of an DNA methylation inhibitor 5-azacytidine (azaC) on the frequency of reversion to a thymidine kinase-positive (TK+) phenotype in 5-bromodexoyuridine (BrdU)-resistant subclones obtained from clones of Chinese hams...