Transitions in replication timing in a 340 kb region of human chromosomal R-Band 1p36.1

• Published in: Journal of cellular biochemistry Vol. 92; no. 4; pp. 755 - 769
• Main Authors: Brylawski, Bruna P., Cohen, Stephanie M., Horne, Hisani, Irani, Neville, Cordeiro-Stone, Marila, Kaufman, David G.
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.07.2004
• Subjects: Aldehyde Dehydrogenase - genetics; chromosome 1; Chromosome Banding; Chromosome Mapping; Chromosomes, Human, Pair 1 - genetics; CpG Islands; DNA Replication - genetics; early S phase; Fibroblasts; Humans; Molecular Weight; Polymerase Chain Reaction; Promoter Regions, Genetic - genetics; quantitative PCR; Replication Origin - genetics; replication origins; replicons; S Phase; Skin; Time Factors
• ISSN: 0730-2312; 1097-4644
• DOI: 10.1002/jcb.20101

DNA replication is initiated within a few chromosomal bands as normal human fibroblasts enter the S phase (Cohen et al. [1998]: Exp Cell Res. 245:321–329). In the present study, we determined the timing of replication of sequences along a 340 kb region in one of these bands, 1p36.13, an R band on chromosome 1. Within this region, we identified a segment of DNA (approximately 140 kb) that is replicated in the first hour of the S phase and is flanked by segments replicated 1–2 h later. Using a quantitative PCR‐based assay to measure sequence abundance in size‐fractionated (900–1,700 nt) nascent DNA (Giacca et al. [1994]: Proc Natl Acad Sci USA. 91:7119), we mapped two functional origins of replication separated by 54 kb and firing 1 h apart. One origin was found to be functional during the first hour of S and was located within a CpG island associated with a predicted gene of unknown function (Genscan NT_004610.2). The second origin was activated in the second hour of S and was mapped to a CpG island near the promoter of the aldehyde dehydrogenase 4A1 (ALDH4A1) gene. At the opposite end of the early replicating segment, a more gradual change in replication timing was observed within the span of approximately 100 kb. These data suggest that DNA replication in adjacent segments of band 1p36.13 is organized differently, perhaps in terms of replicon number and length, or rate of fork progression. In the transition areas that mark the boundaries between different temporal domains, the replication forks initiated in the early replicated region are likely to pause or delay progression before replication of the 340 kb contig is completed. © 2004 Wiley‐Liss, Inc.