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Molecular and Cellular Biology, May 2004, p. 4019-4031, Vol. 24, No. 9
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.9.4019-4031.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Anatomy and Dynamics of DNA Replication Fork Movement in Yeast Telomeric Regions{dagger} ,{ddagger}

Svetlana Makovets, Ira Herskowitz, and Elizabeth H. Blackburn*

University of California, San Francisco, Department of Biochemistry and Biophysics, San Francisco, California 94143-2200

Received 9 December 2003/ Returned for modification 15 January 2004/ Accepted 10 February 2004

Replication initiation and replication fork movement in the subtelomeric and telomeric DNA of native Y' telomeres of yeast were analyzed using two-dimensional gel electrophoresis techniques. Replication origins (ARSs) at internal Y' elements were found to fire in early-mid-S phase, while ARSs at the terminal Y' elements were confirmed to fire late. An unfired Y' ARS, an inserted foreign (bacterial) sequence, and, as previously reported, telomeric DNA each were shown to impose a replication fork pause, and pausing is relieved by the Rrm3p helicase. The pause at telomeric sequence TG1-3 repeats was stronger at the terminal tract than at the internal TG1-3 sequences located between tandem Y' elements. We show that the telomeric replication fork pause associated with the terminal TG1-3 tracts begins ~100 bp upstream of the telomeric repeat tract sequence. Telomeric pause strength was dependent upon telomere length per se and did not require the presence of a variety of factors implicated in telomere metabolism and/or known to cause telomere shortening. The telomeric replication fork pause was specific to yeast telomeric sequence and was independent of the Sir and Rif proteins, major known components of yeast telomeric heterochromatin.

* Corresponding author. Mailing address: Department of Biochemistry and Biophysics, University of California, 600 16th St., San Francisco, San Francisco, CA 94143-2200. Phone: (415) 476-4912. Fax: (415) 514-2913. E-mail: telomer{at}itsa.ucsf.edu.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org.

{ddagger} This paper is dedicated to the memory of Ira Herskowitz (deceased on 28 April 2003).

Molecular and Cellular Biology, May 2004, p. 4019-4031, Vol. 24, No. 9
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.9.4019-4031.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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