Abstract
Human melanoma cells were synchronized with hydroxyurea. The DNA replication intermediates formed either during the hydroxyurea-treatment or after the release of the hydroxyurea-block were investigated. The intermediates were released from the parental DNA by lysing the cells in dilute alkali and were then analyzed by agarose gel electrophoresis. After the release of the hydroxyurea block radioactivity was first detected in Okazaki-fragments and then later also in larger replication intermediates (ranging in size up to 10 kb). The results indicate that the 10 kb DNA replication intermediate is derived from the Okazaki-fragments. Thus during its maturation to high molecular weight DNA the Okazaki-fragments form a large, well defined DNA replication intermediate, which later forms the high molecular weight DNA. However, it is not possible to determine whether all or only a part of the Okazaki-fragments give rise to the 10 kb DNA.
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