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Published online November 19, 2007
doi:10.1083/jcb.200706151
The Journal of Cell Biology, Vol. 179, No. 4, 659-670
The Rockefeller University Press, 0021-9525 $30.00
© 2007 Melloy et al.
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Article

Nuclear fusion during yeast mating occurs by a three-step pathway



Patricia Melloy1,3, Shu Shen1, Erin White2, J. Richard McIntosh2, and Mark D. Rose1

1 Department of Molecular Biology, Princeton University, Princeton, NJ 08544
2 Mollecular, Cellular, and Developmental Biology, University of Colorado at Boulder, Boulder, CO 80309
3 Department of Biological and Allied Health Sciences, Fairleigh Dickinson University, Madison, NJ 07940

Correspondence to Mark D. Rose: mdrose{at}princeton.edu

In Saccharomyces cerevisiae, mating culminates in nuclear fusion to produce a diploid zygote. Two models for nuclear fusion have been proposed: a one-step model in which the outer and inner nuclear membranes and the spindle pole bodies (SPBs) fuse simultaneously and a three-step model in which the three events occur separately. To differentiate between these models, we used electron tomography and time-lapse light microscopy of early stage wild-type zygotes. We observe two distinct SPBs in ~80% of zygotes that contain fused nuclei, whereas we only see fused or partially fused SPBs in zygotes in which the site of nuclear envelope (NE) fusion is already dilated. This demonstrates that SPB fusion occurs after NE fusion. Time-lapse microscopy of zygotes containing fluorescent protein tags that localize to either the NE lumen or the nucleoplasm demonstrates that outer membrane fusion precedes inner membrane fusion. We conclude that nuclear fusion occurs by a three-step pathway.

Abbreviations used in this paper: ET, electron tomography; mRFP, monomeric RFP; NE, nuclear envelope; ROI, region of interest; SPB, spindle pole body; SS, signal sequence.


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