UTILIZATION OF THE SAME DNA-REPLICATION ORIGIN BY HUMAN-CELLS OF DIFFERENT DERIVATION

In the past, a highly sensitive and efficient method was developed to map DNA replication origins in human cells, based on quantitative PCR performed on nascent DNA samples. This method allowed the identificati on of a replication origin in the myeloid HL-60 cell line, located on chromosome 19 within an similar to 500 bp segment near the lamin B2 ge ne [Giacca ct al, (1994) Proc. Natl. Acad. Sci. USA, 91, 7119], The sa me procedure has now been further simplified and extended to a variety of other exponentially growing human cells of different histological derivation (three neural, one connectival and one epithelial), with a nearly diploid chromosomal content. In all the six cell lines tested, the origin activity within the lamin B2 gene domain was localized to t he same region, Furthermore, the lamin B2 origin was also found to be active in stimulated, but not in quiescent, peripheral blood lymphocyt es.