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Molecular and Cellular Biology, January 2002, p. 442-452, Vol. 22, No. 2
0270-7306/01/$04.00+0     DOI: 10.1128/MCB.22.2.442-452.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Replication Initiation Patterns in the ß-Globin Loci of Totipotent and Differentiated Murine Cells: Evidence for Multiple Initiation Regions

Mirit I. Aladjem,1* Luo Wei Rodewald,2 Chii Mai Lin,1 Sarah Bowman,1 Daniel M. Cimbora,3 Linnea L. Brody,2 Elliot M. Epner,4 Mark Groudine,3 and Geoffrey M. Wahl2

Laboratory of Molecular Pharmacology, National Cancer Institute, Bethesda, Maryland 20892-4255,1 Gene Expression Laboratory, The Salk Institute,2 Fred Hutchinson Cancer Research Center, Seattle, Washington 98109,3 Arizona Cancer Center/BMT, Tucson, Arizona 857244

Received 20 August 2001/ Returned for modification 19 September 2001/ Accepted

The replication initiation pattern of the murine ß-globin locus was analyzed in totipotent embryonic stem cells and in differentiated cell lines. Initiation events in the murine ß-globin locus were detected in a region extending from the embryonic Ey gene to the adult ßminor gene, unlike the restricted initiation observed in the human locus. Totipotent and differentiated cells exhibited similar initiation patterns. Deletion of the region between the adult globin genes did not prevent initiation in the remainder of the locus, suggesting that the potential to initiate DNA replication was not contained exclusively within the primary sequence of the deleted region. In addition, a deletion encompassing the six identified 5' hypersensitive sites in the mouse locus control region had no effect on initiation from within the locus. As this deletion also did not affect the chromatin structure of the locus, we propose that the sequences determining both chromatin structure and replication initiation lie outside the hypersensitive sites removed by the deletion.

* Corresponding author. Mailing address: Laboratory of Molecular Pharmacology, DBS/NCI/NIH, Bldg. 37, Rm. 5056D, 37 Convent Dr., Bethesda, MD 20892-4255. Phone: (301) 435-2848. Fax: (301) 402-0752. E-mail: maladjem{at}helix.nih.gov.

Molecular and Cellular Biology, January 2002, p. 442-452, Vol. 22, No. 2
0022-538X/01/$04.00+0     DOI: 10.1128/MCB.22.2.442-452.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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