Allele-specific replication timing in imprinted domains: absence of asynchrony at several loci.
Using a bromodeoxyuridine incorporation method to detect replicated DNA, we studied allele-specific replication of several sites within the human Prader-Willi/Angelman and IGF2/H19 imprinted regions. No obvious allele-specific differences in time of replication were detected at most loci previously reported to replicate asynchronously in the same cell types as determined by a FISH-based replication assay. Our finding of an absence of allelic replication asynchrony may be related to low levels of imprinted gene expression near these loci in the examined cells (lymphocytes, fibroblasts and lymphoblastoid cells). This view is supported by our studies of the imprinted SNRPN gene in that cells with paternal allele-specific expression (lymphocytes and lymphoblasts) replicate SNRPN alleles asynchronously, whereas cells with a low level of expression (HeLa) replicate SNRPN later and with less allelic asynchrony. In lymphoblasts, the early replicating allele of SNRPN was identified as the paternal one based on the properties of maternal allele-specific methylation and paternal allele-specific expression. Our studies suggest that FISH data implying replication asynchrony in nonexpressing cells reflect structural differences between the maternal and paternal alleles rather than differences in replication timing.
Kawame
H
H
Department of Medicine, University of Washington, Seattle 98195, USA.
Gartler
S M
SM
Hansen
R S
RS
D000483:Alleles; D001324:Autoantigens / Q000235:genetics; D001483:Base Sequence; D001973:Bromodeoxyuridine; D002461:Cell Line, Transformed; D004247:DNA; D004261:DNA Replication; D018392:Genomic Imprinting; D006367:HeLa Cells; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007335:Insulin-Like Growth Factor II / Q000235:genetics; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000235:genetics; D011218:Prader-Willi Syndrome / Q000235:genetics; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated; D017411:Ribonucleoproteins, Small Nuclear; D016324:Sequence Tagged Sites; D013997:Time Factors; D055517:snRNP Core Proteins
D001324:Autoantigens; C094335:H19 long non-coding RNA; D009124:Muscle Proteins; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated; D017411:Ribonucleoproteins, Small Nuclear; C526417:SNRPN protein, human; D055517:snRNP Core Proteins; D007335:Insulin-Like Growth Factor II; D004247:DNA; D001973:Bromodeoxyuridine