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Molecular and Cellular Biology, June 2002, p. 4202-4217, Vol. 22, No. 12
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.12.4202-4217.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Dynamic Localization of an Okazaki Fragment Processing Protein Suggests a Novel Role in Telomere Replication

Wonchae Choe,1 Martin Budd,1 Osamu Imamura,1 Laura Hoopes,2 and Judith L. Campbell1*

Braun Laboratories, Pasadena, California 91125,1 Department of Biology and Molecular Biology Program, Pomona College, Claremont, California 917112

Received 19 November 2001/ Returned for modification 2 January 2002/ Accepted 22 February 2002

We have found that the Dna2 helicase-nuclease, thought to be involved in maturation of Okazaki fragments, is a component of telomeric chromatin. We demonstrate a dynamic localization of Dna2p to telomeres that suggests a dual role for Dna2p, one in telomere replication and another, unknown function, perhaps in telomere capping. Both chromatin immunoprecipitation (ChIP) and immunofluorescence show that Dna2p associates with telomeres but not bulk chromosomal DNA in G1 phase, when there is no telomere replication and the telomere is transcriptionally silenced. In S phase, there is a dramatic redistribution of Dna2p from telomeres to sites throughout the replicating chromosomes. Dna2p is again localized to telomeres in late S, where it remains through G2 and until the next S phase. Telomeric localization of Dna2p required Sir3p, since the amount of Dna2p found at telomeres by two different assays, one-hybrid and ChIP, is severely reduced in strains lacking Sir3p. The Dna2p is also distributed throughout the nucleus in cells growing in the presence of double-strand-break-inducing agents such as bleomycin. Finally, we show that Dna2p is functionally required for telomerase-dependent de novo telomere synthesis and also participates in telomere lengthening in mutants lacking telomerase.

* Corresponding author. Mailing address: Braun Laboratories, 147-75, Pasadena, CA 91125. Phone: (626) 395-6053. Fax: (626) 405-9452. E-mail: jcampbel{at}cco.caltech.edu.

Molecular and Cellular Biology, June 2002, p. 4202-4217, Vol. 22, No. 12
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.12.4202-4217.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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