10 kb DNA replication intermediates are formed from Okazaki-fragments in human malignant melanoma cells |
| |
| Affiliation: | 1. Department of Laboratory Medicine and Pathobiology, Temerty Faculty of Medicine, University of Toronto, MaRS Centre, West Tower, 661 University Avenue, Toronto, ON M5G 1M1, Canada;2. Canada Research Chairs Program, Temerty Faculty of Medicine, University of Toronto, Medical Sciences Building, 1 King’s College Circle, Toronto, ON M5S 1A8, Canada;1. Department of Biochemistry, Boston University School of Medicine, Boston, MA, USA |
| |
| 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. |
| |
| Keywords: | |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
