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Title: Phenotypic switching in cells transformed with the herpes simplex virus thymidine kinase gene

Abstract

Biochemical transformation of Ltk/sup -/ cells with the herpes simplex virus thymidine kinase (tk) gene resulted in numerous TK/sup +/ colonies that survived selection in hypoxanthine-aminopterin-thymidine medium. Many of these TK/sup +/ cell lines switched phenotypes and reverted to the TK/sup -/ state. In this report, the authors describe the biological and biochemical characteristics of three TK/sup -/ revertant lines. One (K/sub 1/B/sub 5/) transiently expressed TK in the presence of bromodeoxyuridine, which selects for the TK/sup -/ phenotype. Another TK/sup -/ sibling (K/sub 1/B/sub 6//sup n/) expressed TK only after removal from bromodeoxyuridine-containing medium. The last variant (K/sub 1/B/sub 6//sup me/) lost the ability to switch to the TK/sup +/ phenotype, although it maintained the herpes simplex virus sequences coding for TK. Loss of the ability of K/sub 1/B/sub 6//sup me/ 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 nomore » longer methylated.« less

Authors:
; ;
Publication Date:
Research Org.:
Dept. of Microbiology and The Cancer Research Center, College of Physicians and Surgeons, Columbia Univ., New York, NY 10032
OSTI Identifier:
5809822
Resource Type:
Journal Article
Journal Name:
Mol. Cell. Biol.; (United States)
Additional Journal Information:
Journal Volume: 2:6
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; BUDR; TOXICITY; ENDONUCLEASES; ENZYME INHIBITORS; HERPES SIMPLEX; DNA SEQUENCING; PHOSPHOTRANSFERASES; BIOLOGICAL FUNCTIONS; GENES; REVERTANTS; BIOCHEMICAL REACTION KINETICS; CELL CULTURES; CELL TRANSFORMATIONS; COLONY FORMATION; CULTURE MEDIA; HYPOXANTHINE; MESSENGER-RNA; METHYLATION; MUTAGENESIS; PHENOTYPE; THYMIDINE; ANTIMETABOLITES; AROMATICS; AZAARENES; AZINES; BROMOURACILS; CHEMICAL REACTIONS; DISEASES; DNA-ASE; DRUGS; ENZYMES; ESTERASES; FUNCTIONS; HETEROCYCLIC COMPOUNDS; HYDROLASES; HYDROXY COMPOUNDS; INFECTIOUS DISEASES; KINETICS; MUTANTS; NUCLEIC ACIDS; NUCLEOSIDES; NUCLEOTIDES; ORGANIC BROMINE COMPOUNDS; ORGANIC COMPOUNDS; ORGANIC HALOGEN COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; PHOSPHODIESTERASES; PHOSPHORUS-GROUP TRANSFERASES; PURINES; PYRIMIDINES; REACTION KINETICS; RIBOSIDES; RNA; SKIN DISEASES; STRUCTURAL CHEMICAL ANALYSIS; TRANSFERASES; URACILS; VIRAL DISEASES; 550400* - Genetics; 550700 - Microbiology

Citation Formats

Ostrander, M, Vogel, S, and Silverstein, S. Phenotypic switching in cells transformed with the herpes simplex virus thymidine kinase gene. United States: N. p., 1982. Web. doi:10.1128/MCB.2.6.708.
Ostrander, M, Vogel, S, & Silverstein, S. Phenotypic switching in cells transformed with the herpes simplex virus thymidine kinase gene. United States. https://doi.org/10.1128/MCB.2.6.708
Ostrander, M, Vogel, S, and Silverstein, S. Tue . "Phenotypic switching in cells transformed with the herpes simplex virus thymidine kinase gene". United States. https://doi.org/10.1128/MCB.2.6.708.
@article{osti_5809822,
title = {Phenotypic switching in cells transformed with the herpes simplex virus thymidine kinase gene},
author = {Ostrander, M and Vogel, S and Silverstein, S},
abstractNote = {Biochemical transformation of Ltk/sup -/ cells with the herpes simplex virus thymidine kinase (tk) gene resulted in numerous TK/sup +/ colonies that survived selection in hypoxanthine-aminopterin-thymidine medium. Many of these TK/sup +/ cell lines switched phenotypes and reverted to the TK/sup -/ state. In this report, the authors describe the biological and biochemical characteristics of three TK/sup -/ revertant lines. One (K/sub 1/B/sub 5/) transiently expressed TK in the presence of bromodeoxyuridine, which selects for the TK/sup -/ phenotype. Another TK/sup -/ sibling (K/sub 1/B/sub 6//sup n/) expressed TK only after removal from bromodeoxyuridine-containing medium. The last variant (K/sub 1/B/sub 6//sup me/) lost the ability to switch to the TK/sup +/ phenotype, although it maintained the herpes simplex virus sequences coding for TK. Loss of the ability of K/sub 1/B/sub 6//sup me/ 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.},
doi = {10.1128/MCB.2.6.708},
url = {https://www.osti.gov/biblio/5809822}, journal = {Mol. Cell. Biol.; (United States)},
number = ,
volume = 2:6,
place = {United States},
year = {1982},
month = {6}
}