title: ER-associated SNAREs and Sey1p mediate nuclear fusion at two distinct steps during yeast mating.
abstract: During yeast mating, two haploid nuclei fuse membranes to form a single diploid nucleus. However, the known proteins required for nuclear fusion are unlikely to function as direct fusogens (i.e., they are unlikely to directly catalyze lipid bilayer fusion) based on their predicted structure and localization. Therefore we screened known fusogens from vesicle trafficking (soluble N-ethylmaleimide-sensitive factor attachment protein receptors [SNAREs]) and homotypic endoplasmic reticulum (ER) fusion (Sey1p) for additional roles in nuclear fusion. Here we demonstrate that the ER-localized SNAREs Sec20p, Ufe1p, Use1p, and Bos1p are required for efficient nuclear fusion. In contrast, Sey1p is required indirectly for nuclear fusion; sey1Δ zygotes accumulate ER at the zone of cell fusion, causing a block in nuclear congression. However, double mutants of Sey1p and Sec20p, Ufe1p, or Use1p, but not Bos1p, display extreme ER morphology defects, worse than either single mutant, suggesting that retrograde SNAREs fuse ER in the absence of Sey1p. Together these data demonstrate that SNAREs mediate nuclear fusion, ER fusion after cell fusion is necessary to complete nuclear congression, and there exists a SNARE-mediated, Sey1p-independent ER fusion pathway.
authors.affiliation: Department of Molecular Biology, Princeton University, Princeton, NJ 08544-1014.
authors.forename: Jason V
authors.initials: JV
authors.lastname: Rogers
authors.forename: Tim
authors.initials: T
authors.lastname: Arlow
authors.forename: Elizabeth R
authors.initials: ER
authors.lastname: Inkellis
authors.forename: Timothy S
authors.initials: TS
authors.lastname: Koo
authors.forename: Mark D
authors.initials: MD
authors.lastname: Rose
mesh_terms: D002467:Cell Nucleus / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D008561:Membrane Fusion; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D050765:Qa-SNARE Proteins / Q000235:genetics / Q000378:metabolism; D050766:Qb-SNARE Proteins / Q000235:genetics / Q000378:metabolism; D050767:Qc-SNARE Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D033921:Vesicular Transport Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: C068037:BOS1 protein, S cerevisiae; D008562:Membrane Glycoproteins; D050765:Qa-SNARE Proteins; D050766:Qb-SNARE Proteins; D050767:Qc-SNARE Proteins; C073173:SEC20 protein, S cerevisiae; C472231:SEY1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C103868:UFE1 protein, S cerevisiae; C477343:Use1 protein, S cerevisiae; D033921:Vesicular Transport Proteins