title: Asynchronous replication timing of imprinted loci is independent of DNA methylation, but consistent with differential subnuclear localization.
abstract: Genomic imprinting in mammals marks the two parental alleles resulting in differential gene expression. Imprinted loci are characterized by distinct epigenetic modifications such as differential DNA methylation and asynchronous replication timing. To determine the role of DNA methylation in replication timing of imprinted loci, we analyzed replication timing in Dnmt1- and Dnmt3L-deficient embryonic stem (ES) cells, which lack differential DNA methylation and imprinted gene expression. Asynchronous replication is maintained in these ES cells, indicating that asynchronous replication is parent-specific without the requirement for differential DNA methylation. Imprinting centers are required for regional control of imprinted gene expression. Analysis of replication fork movement and three-dimensional RNA and DNA fluorescent in situ hybridization (FISH) analysis of the Igf2-H19 locus in various cell types indicate that the Igf2-H19 imprinting center differentially regulates replication timing of nearby replicons and subnuclear localization. Based on these observations, we suggest a model in which cis elements containing nonmethylation imprints are responsible for the movement of parental imprinted loci to distinct nuclear compartments with different replication characteristics resulting in asynchronous replication timing.
authors.affiliation: Whitehead Institute for Biomedical Research, Cambridge, MA 02142, USA.
authors.forename: Joost
authors.initials: J
authors.lastname: Gribnau
authors.forename: Konrad
authors.initials: K
authors.lastname: Hochedlinger
authors.forename: Ken
authors.initials: K
authors.lastname: Hata
authors.forename: En
authors.initials: E
authors.lastname: Li
authors.forename: Rudolf
authors.initials: R
authors.lastname: Jaenisch
mesh_terms: D000483:Alleles; D000818:Animals; D001973:Bromodeoxyuridine / Q000494:pharmacology; D002467:Cell Nucleus / Q000378:metabolism; D019175:DNA Methylation; D005260:Female; D005434:Flow Cytometry; D018392:Genomic Imprinting; D017404:In Situ Hybridization, Fluorescence; D007335:Insulin-Like Growth Factor II / Q000378:metabolism; D008297:Male; D051379:Mice; D008898:Mimosine / Q000494:pharmacology; D008957:Models, Genetic; D009154:Mutation; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated / Q000378:metabolism; D016196:S Phase; D013347:Subcellular Fractions; D013997:Time Factors
chemical_list: C094335:H19 long non-coding RNA; D016376:Oligonucleotides, Antisense; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated; D008898:Mimosine; D007335:Insulin-Like Growth Factor II; D001973:Bromodeoxyuridine