Increased origin activity in transformed versus normal cells: identification of novel protein players involved in DNA replication and cellular transformation.

ABSTRACT: Using libraries of replication origins generated previously, we identified three clones that supported the autonomous replication of their respective plasmids in transformed, but not in normal cells. Assessment of their in vivo replication activity by in situ chromosomal DNA replication assays revealed that the chromosomal loci corresponding to these clones coincided with chromosomal replication origins in all cell lines, which were more active by 2-3-fold in the transformed by comparison to the normal cells. Evaluation of pre-replication complex (pre-RC) protein abundance at these origins in transformed and normal cells by chromatin immunoprecipitation assays, using anti-ORC2, -cdc6 and -cdt1 antibodies, showed that they were bound by these pre-RC proteins in all cell lines, but a 2-3-fol

SUBMITTER: Di Paola D 

PROVIDER: S-EPMC2853114 | biostudies-literature | 2010 Apr

REPOSITORIES: biostudies-literature