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Molecular and Cellular Biology, January 2005, p. 303-311, Vol. 25, No. 1
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.1.303-311.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Molecular and Cellular Dissection of Mating-Type Switching Steps in Schizosaccharomyces pombe

Allyson M. Holmes, Atanas Kaykov, and Benoit Arcangioli*

Unité de Dynamique du Génome, URA 1644 du CNRS, Departement de la Structure et Dynamique des Génomes, Institut Pasteur, Paris, France

Received 26 July 2004/ Returned for modification 20 September 2004/ Accepted 4 October 2004

A strand-specific imprint (break) controls mating-type switching in fission yeast. By introducing a thiamine repressible promoter upstream of the mat1 locus, we can force transcription through the imprinted region, erasing the imprint and inhibiting further mating-type switching, in a reversible manner. Starting from a synchronized, virgin M-cell population, we show that the site- and strand-specific break is formed when DNA replication intermediates appear at mat1 during the first S phase. The formation of the break is concomitant with a replication fork pause and binding of the Swi1 protein at mat1 until early G2 and then rapidly disappears. Upon its formation, the break remains stable throughout the cell cycle and triggers mating-type switching during the second S phase. Finally, we have recreated the mating-type switching pedigree at the molecular and single-cell levels, allowing for the first time separation between the establishment of imprinting and its developmental fate.

* Corresponding author. Mailing address: Unité de Dynamique du Génome, URA 1644 du CNRS, Departement de la Structure et Dynamique des Génomes, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris Cedex 15, France. Phone: 33 1 4568 8454. Fax: 33 1 4568 8960. E-mail: barcan{at}pasteur.fr.

Molecular and Cellular Biology, January 2005, p. 303-311, Vol. 25, No. 1
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.1.303-311.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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