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Published online 26 January 2004. doi:10.1083/jcb.200311063
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 164, Number 3, 361-371
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Article

 Mating type–dependent constraints on the mobility of the left arm of yeast chromosome III

Debra A. Bressan1, Julio Vazquez2, and James E. Haber1

1 Rosenstiel Center and Department of Biology, Brandeis University, Waltham, MA 02454
2 Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA 94143

Address correspondence to James E. Haber, Rosenstiel Center and Dept. of Biology, Brandeis University, Waltham, MA 02454-9910. Tel.: (781) 736-2462. Fax: (781) 736-2405. email: haber{at}brandeis.edu

Mating-type gene (MAT) switching in budding yeast exhibits donor preference. MATa preferentially recombines with HML near the left telomere of chromosome III, whereas MAT{alpha} prefers HMR near the right telomere. Donor preference is controlled by the recombination enhancer (RE) located proximal to HML. To test if HML is constrained in pairing with MAT{alpha}, we examined live-cell mobility of LacI-GFP–bound lactose operator (lacO) arrays inserted at different chromosomal sites. Without induction of recombination, lacO sequences adjacent to HML are strongly constrained in both MAT{alpha} and RE-deleted MATa strains, compared with MATa. In contrast, chromosome movement at HMR or near a telomere of chromosome V is mating-type independent. HML is more constrained in MATa {Delta}re and less constrained in MATa RE+ compared with other sites. Although HML and MATa are not prealigned before inducing recombination, the three-dimensional configuration of MAT, HML, and HMR is mating-type dependent. These data suggest there is constitutive tethering of HML, which is relieved in MATa cells through the action of RE.

Key Words: Saccharomyces cerevisiae; mating-type switching; donor preference; deconvolution fluorescence microscopy; chromosome dynamics

Julio Vazquez's present address is Fred Hutchinson Cancer Research Center, P.O. Box 19024, DE-512, Seattle, WA 98109-1024.

Abbreviations used in this paper: 3D, three-dimensional; DSB, double-strand break; LacI, lactose repressor; lacO, lactose operator; MAT, mating-type gene; MSD, mean-squared change in distance; RE, recombination enhancer; tetO, tetracycline operator; TetR, tetracycline repressor; YPD, yeast extract-peptone-dextrose.


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