High-throughput mapping of origins of replication in human cells.
Mapping origins of replication has been challenging in higher eukaryotes. We have developed a rapid, genome-wide method to map origins of replication in asynchronous human cells by combining the nascent strand abundance assay with a highly tiled microarray platform, and we validated the technique by two independent assays. We applied this method to analyse the enrichment of nascent DNA in three 50-kb regions containing known origins of replication in the MYC, lamin B2 (LMNB2) and haemoglobin beta (HBB) genes, a 200-kb region containing the rare fragile site, FRAXA, and a 1,075-kb region on chromosome 22; we detected most of the known origins and also 28 new origins. Surprisingly, the 28 new origins were small in size and located predominantly within genes. Our study also showed a strong correlation between origin replication timing and chromatin acetylation.
Department of Medicine, Section of Hematology/Oncology, The University of Chicago, Chicago, Illinois 60637, USA.
Isabelle
I
Lucas
Aparna
A
Palakodeti
Yanwen
Y
Jiang
David J
DJ
Young
Nan
N
Jiang
Anthony A
AA
Fernald
Michelle M
MM
Le Beau
D000107:Acetylation; D002478:Cells, Cultured; D002843:Chromatin / Q000378:metabolism; D047369:Chromatin Immunoprecipitation; D004261:DNA Replication / Q000235:genetics*; D015894:Genome, Human / Q000235:genetics*; D006801:Humans; D020161:Physical Chromosome Mapping / Q000379:methods*; D018741:Replication Origin / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D002843:Chromatin