Viral and Cellular DNA Synthesis in Nuclei from Human Lymphocytes Transformed by Epstein-Barr Virus

Viral and Cellular DNA Synthesis in Nuclei from Human Lymphocytes Transformed by Epstein-Barr Virus

A DNA-synthesizing system in vitro, using nuclei prepared by treatment of human lymphocytes with the detergent Brij 58, was developed. Nuclei from cultured lymphocytes synthesized DNA for as long as 5 hr, and required ATP, deoxynucleoside triphosphates, magnesium, and a calcium chelator. In nuclei f...

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Veröffentlicht in:Proceedings of the National Academy of Sciences - PNAS 1975-06, Vol.72 (6), p.2413-2417
Hauptverfasser: Benz, Wendy C., Strominger, Jack L.
Format: Artikel
Sprache:eng
Schlagworte:
Adenosine Triphosphate - pharmacology
Bromodeoxyuridine - metabolism
Cell Fractionation
Cell Line
Cell lines
Cell nucleus
Cell Nucleus - drug effects
Cell Nucleus - metabolism
Centrifugation, Density Gradient
Detergents
Detergents - pharmacology
DNA
DNA - biosynthesis
DNA, Viral - biosynthesis
Egtazic Acid - pharmacology
Herpesvirus 4, Human
Human herpesvirus 4
Humans
Interphase
Kinetics
Lectins - pharmacology
Lymphocyte Activation
Lymphocytes
Lymphocytes - drug effects
Lymphocytes - metabolism
Lymphocytes - ultrastructure
Magnesium - pharmacology
Radioactive decay
T lymphocytes
Transformed cell line
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Zusammenfassung:A DNA-synthesizing system in vitro, using nuclei prepared by treatment of human lymphocytes with the detergent Brij 58, was developed. Nuclei from cultured lymphocytes synthesized DNA for as long as 5 hr, and required ATP, deoxynucleoside triphosphates, magnesium, and a calcium chelator. In nuclei from a partially synchronized line of cultured lymphocytes carrying several hundred copies of the Epstein-Barr viral genome, synthesis in vitro was predominantly viral in early S phase and cellular in late S phase. These and other data suggested that the DNA synthesis observed in vitro was predominantly replicative.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.72.6.2413