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Eukaryotic Cell, October 2009, p. 1554-1566, Vol. 8, No. 10
1535-9778/09/$08.00+0     doi:10.1128/EC.00209-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Aneuploid Chromosomes Are Highly Unstable during DNA Transformation of Candida albicans{triangledown}

Kelly Bouchonville,1 Anja Forche,1,2 Karen E. S. Tang,1 Anna Selmecki,1,{dagger} and Judith Berman1,3*

Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota 55455,1 Department of Biology, Bowdoin College, Brunswick, Maine 04011,2 Department of Microbiology, University of Minnesota, Minneapolis, Minnesota 554553

Received 17 July 2009/ Accepted 16 August 2009

Candida albicans strains tolerate aneuploidy, historically detected as karyotype alterations by pulsed-field gel electrophoresis and more recently revealed by array comparative genome hybridization, which provides a comprehensive and detailed description of gene copy number. Here, we first retrospectively analyzed 411 expression array experiments to predict the frequency of aneuploidy in different strains. As expected, significant levels of aneuploidy were seen in strains exposed to stress conditions, including UV light and/or sorbose treatment, as well as in strains that are resistant to antifungal drugs. More surprisingly, strains that underwent transformation with DNA displayed the highest frequency of chromosome copy number changes, with strains that were initially aneuploid exhibiting ~3-fold more copy number changes than strains that were initially diploid. We then prospectively analyzed the effect of lithium acetate (LiOAc) transformation protocols on the stability of trisomic chromosomes. Consistent with the retrospective analysis, the proportion of karyotype changes was highly elevated in strains carrying aneuploid chromosomes. We then tested the hypothesis that stresses conferred by heat and/or LiOAc exposure promote chromosome number changes during DNA transformation procedures. Indeed, a short pulse of very high temperature caused frequent gains and losses of multiple chromosomes or chromosome segments. Furthermore, milder heat exposure over longer periods caused increased levels of loss of heterozygosity. Nonetheless, aneuploid chromosomes were also unstable when strains were transformed by electroporation, which does not include a heat shock step. Thus, aneuploid strains are particularly prone to undergo changes in chromosome number during the stresses of DNA transformation protocols.

* Corresponding author. Mailing address: University of Minnesota, GCD, 6-160 Jackson Hall, 321 Church St. SE, Minneapolis, MN 55455. Phone: (612) 625-1971. Fax: (612) 625-5754. E-mail: jberman{at}umn.edu

{triangledown} Published ahead of print on 21 August 2009.

{dagger} Present address: Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, MA 02115.

Eukaryotic Cell, October 2009, p. 1554-1566, Vol. 8, No. 10
1535-9778/09/$08.00+0     doi:10.1128/EC.00209-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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