Phenotypic Switching in Cells Transformed with the Herpes Simplex Virus Thymidine Kinase Gene
AUTOR(ES)
Ostrander, Michael
RESUMO
Biochemical transformation of Ltk− cells with the herpes simplex virus thymidine kinase (tk) gene resulted in numerous TK+ colonies that survived selection in hypoxanthine-aminopterin-thymidine medium. Many of these TK+ cells lines switched phenotypes and reverted to the TK− state. In this report, we describe the biological and biochemical characteristics of three TK− revertant lines. One (K1B5) transiently expressed TK in the presence of bromodeoxyuridine, which selects for the TK− phenotype. Another TK− sibling (K1B6n) expressed TK only after removal from bromodeoxyuridine-containing medium. The last variant (K1B6me) lost the ability to switch to the TK+ phenotype, although it maintained the herpes simplex virus sequences coding for TK. Loss of the ability of K1B6me cells to express TK was correlated with extensive methylation of the sequence recognized by the restriction endonuclease HpaII (pCpCpGpG). After these cells were treated with 5-azacytidine, they regained the ability to clone in hypoxanthine-aminopterin-thymidine medium and reexpressed virus tk mRNA and enzyme. In addition, the HpaII sites that were previously shown to be refractile to enzyme digestion were converted to a sensitive state, demonstrating that they were no longer methylated.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=369847Documentos Relacionados
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