Two Epstein-Barr virus-associated DNA polymerase activities.

Epstein-Barr virus induces a unique pyrimidine deoxynucleoside kinase activity in superinfected and virus-producer B cell lines

“…In the case of EB virus-induced DNA polymerase and nuclease, it has been shown that nonproducer Raji cells contain only host enzymatic activities and that virus-induced enzymes appear in Raji cells only after EB virus superinfection or drug treatment (Goodman et al, 1978;Clough, 1979;Feighny et al, 1980;Ooka & Calender, 1980;Clough & McMahon, 1981). Also, EB virus-induced enzymes are found in viral capsid antigen-positive producer cell populations, namely, HR-1 and B95-8 cell lines (Goodman et al, 1978;Clough, 1979). Therefore, we postulated that any EB virus-induced pyrimidine deoxynucleoside kinase activity would be induced in Raji cells by superinfection and would be present in EB virus-producer cell lines.…”

“…Biochemical studies were performed on those fractions exhibiting dTk or dCk activities obtained from DEAE-cellulose chromatograms of HR-1 cell extracts. In spite of the much smaller amount of EB virusinduced dTk activity present in HR-1 as compared to B95-8 producer cells, pyrimidine deoxynucleoside kinase activities from HR-1 cells were used for these characterization experiments because (1) HR-1 virus was used in the Raji superinfection experiments shown in Figures 1 and 2 and (2) characterization of other EB virus-induced enzymes was performed with material isolated from HR-1 cells (Goodman et al, 1978;Clough, 1979Clough, , 1980. Fractions containing the dTk activity eluting with 20 mM K2P04 in the void volume were pooled and considered to represent cellular or cytosol dTk.…”

“…DEAE-cellulose Column Chromatography. Procedures for harvesting cells, making crude extracts, and performing DEAE-cellulose column chromatography have been previously described by Goodman et al (1978) andClough (1979).…”

“…Herpes viruses induce most of the enzymes required for replication of viral DNA in productively infected cells. These viral enzymes include DNA polymerase (Weissbach et al, 1973;Boezi et al, 1974;Huang, 1975;Allen et al, 1977;Goodman et al, 1978), DNase (Hoffman & Cheng, 1978;Clough, 1979Clough, ,1980; Hoffman, 1981), and a recently reported topoisomerase (Biswal et al, 1983). Virus-induced pyrimidine deoxynucleoside kinase [thymidine kinase (dTk)]1 activity has tThis work was supported by National Institutes of Health Grant CA23070 and Research Career Development Award CA00707.…”

“…figure 3: DEAE-cellulose chromatography of cell extracts fromvarious cell lines. Extracts were prepared as described under Experimental Procedures and byGoodman et al (1978) andClough (1979) and passed through DEAE-cellulose columns, and alternate fractions were tested for thymidine kinase (•) or deoxycytidine kinase (O) activity. (A) Cell extract from 1.0 L of HR-1 cells; (B) cell extract from 1.0 L of B95-8 cejls; (C) cell extract from 1.0 L of Raji cells.…”