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Complementation of replication origin function in mouse embryonic stem cells by human DNA sequences.

Stephanie M Cohen·Seigo Hatada·Bruna P Brylawski·Oliver Smithies·David G Kaufman·Marila Cordeiro-Stone
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Abstract

A functional origin of replication was mapped to the transcriptional promoter and exon 1 of the hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene in the mouse and human genomes. This origin was lost in mouse embryonic stem (ES) cells with a spontaneous deletion (approximately 36 kb) at the 5' end of the HPRT locus. Restoration of HPRT activity by homologous recombination with human/mouse chimeric sequences reconstituted replication origin activity in two independent ES cell lines. Quantitative PCR analyses of abundance of genetic markers in size-fractionated nascent DNA indicated that initiation of DNA replication coincided with the site of insertion in the mouse genome of the 335 bp of human DNA containing the HPRT exon 1 and a truncated promoter. The genetic information contained in the human sequence and surrounding mouse DNA was analyzed for cis-acting elements that might contribute to selection and functional activation of a mammalian origin of DNA replication.

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AnimalsBase SequenceDNA PrimersGene ComponentsHumansHypoxanthine PhosphoribosyltransferaseMiceMolecular Sequence DataPolymerase Chain ReactionRecombination, GeneticReplication OriginSequence AlignmentStem CellsComparative StudyJournal ArticleResearch Support, U.S. Gov't, P.H.S.

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Stephanie M Cohen, Seigo Hatada, Bruna P Brylawski, Oliver Smithies, David G Kaufman, Marila Cordeiro-Stone. 2004. Complementation of replication origin function in mouse embryonic stem cells by human DNA sequences.. https://doi.org/10.1016/j.ygeno.2004.04.008

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