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Cells of the K562 human erythroleukemia cell line were obtained in different stages of the cell cycle by centrifugal elutriation. The cells had been previously labeled for 2 hr with BrdUrd so that BrdUrd-DNA synthesized during four different selected intervals of the S phase could be isolated. This DNA was used to determine the temporal replication during S phase of EcoRI segments containing histone genes. Cloned human genomic segments containing the core histone genes (H2A, H2B, H3, and H4), H2A and H2B pseudogenes, and the H1 gene were prepared. The genomic inserts were excised from these plasmids, nick-translated, and used as hybridization probes. The results with different probes compared on the same and on independently prepared DBM-paper transfers indicate that all of these histone genes replicate during the first half of the S phase. These genes were not among the earliest to replicate in the K562 cell line. Similar studies were carried out with HeLa cells in which EcoRI segments containing the H4 histone and H2A and H2B pseudogenes were found to replicate during the first half of the S phase. These histone genes replicate during the interval of the S phase when histone mRNA appears in the cytoplasm at the maximal rate. The possible relationship between these events is discussed.

Coordinate replication of members of the multigene family of core and H1 human histone genes.