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Molecular and Cellular Biology, May 2004, p. 4138-4150, Vol. 24, No. 10
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.10.4138-4150.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Defined Sequence Modules and an Architectural Element Cooperate To Promote Initiation at an Ectopic Mammalian Chromosomal Replication Origin

Amy L. Altman{dagger} and Ellen Fanning*

Department of Biological Sciences and the Vanderbilt-Ingram Cancer Center, Vanderbilt University, Nashville, Tennessee 37235

Received 27 June 2003/ Returned for modification 12 August 2003/ Accepted 19 February 2004

A small DNA fragment containing the high-frequency initiation region (IR) ori-ß from the hamster dihydrofolate reductase locus functions as an independent replicator in ectopic locations in both hamster and human cells. Conversely, a fragment of the human lamin B2 locus containing the previously mapped IR serves as an independent replicator at ectopic chromosomal sites in hamster cells. At least four defined sequence elements are specifically required for full activity of ectopic ori-ß in hamster cells. These include an AT-rich element, a 4-bp sequence located within the mapped IR, a region of intrinsically bent DNA located between these two elements, and a RIP60 protein binding site adjacent to the bent region. The ori-ß AT-rich element is critical for initiation activity in human, as well as hamster, cells and can be functionally substituted for by an AT-rich region from the human lamin B2 IR that differs in nucleotide sequence and length. Taken together, the results demonstrate that two mammalian replicators can be activated at ectopic sites in chromosomes of another mammal and lead us to speculate that they may share functionally related elements.

* Corresponding author. Mailing address: Department of Biological Sciences, Vanderbilt University, 2325 Stevenson Center, 1161 21st Ave. S., Nashville, TN 37235-1634. Phone: (615) 343-5677. Fax: (615) 343-6707. E-mail: ellen.h.fanning{at}vanderbilt.edu.

{dagger} Present address: Booz Allen Hamilton, Inc., Falls Church, VA 22042.

Molecular and Cellular Biology, May 2004, p. 4138-4150, Vol. 24, No. 10
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.10.4138-4150.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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