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Vol. 14, Issue 6, 2530-2542, June 2003

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The Chlamydomonas Fus1 Protein Is Present on the Mating Type plus Fusion Organelle and Required for a Critical Membrane Adhesion Event during Fusion with minus Gametes

Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9039

Submitted December 12, 2002; Revised January 29, 2003; Accepted February 26, 2003
Monitoring Editor: Paul M. Wassarman

The molecular mechanisms of the defining event in fertilization, gamete fusion, remain poorly understood. The FUS1 gene in the unicellular, biflagellated green alga Chlamydomonas is one of the few sex-specific eukaryotic genes shown by genetic analysis to be essential for gamete fusion during fertilization. In Chlamydomonas, adhesion and fusion of the plasma membranes of activated mt+ and mt- gametes is accomplished via specialized fusion organelles called mating structures. Herein, we identify the endogenous Fus1 protein, test the idea that Fus1 is at the site of fusion, and identify the step in fusion that requires Fus1. Our results show that Fus1 is a 95-kDa protein present on the external surface of both unactivated and activated mt+ gametes. Bioassays indicate that adhesion between mating type plus and mating type minus fusion organelles requires Fus1 and that Fus1 is functional only after gamete activation. Finally, immunofluorescence demonstrates that the Fus1 protein is present as an apical patch on unactivated gametes and redistributes during gamete activation over the entire surface of the microvillous-like activated plus mating structure, the fertilization tubule. Thus, Fus1 is present on mt+ gametes at the site of cell-cell fusion and essential for an early step in the fusion process.

^* Corresponding author. E-mail address: william.snell{at}UTSouthwestern.edu.

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