Elion E A, Trueheart J, Fink G R
Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA.
J Cell Biol. 1995 Sep;130(6):1283-96. doi: 10.1083/jcb.130.6.1283.
Zygote formation occurs through tightly coordinated cell and nuclear fusion events. Genetic evidence suggests that the FUS2 gene product promotes cell fusion during zygote formation in Saccharomyces cerevisiae, functioning with the Fus1 plasma membrane protein at or before cell wall and plasma membrane fusion. Here we report the sequence of the FUS2 gene, localization of Fus2 protein, and show that fus1 and fus2 mutants have distinct defects in cell fusion. FUS2 encodes a unique open reading frame of 617 residues that only is expressed in haploid cells in response to mating pheromone. Consistent with a role in cell fusion, Fus2 protein localizes with discrete structures that could be of cytoskeletal or vesicular origin that accumulate at the tip of pheromone-induced shmoos and at the junction of paired cells in zygotes. Fus2 is predicted to be a coiled-coil protein and fractionates with a 100,000 g pellet, suggesting that it is associated with cytoskeleton, membranes, or other macromolecular structures. Fus2 may interact with structures involved in the alignment of the nuclei during cell fusion, because fus2 mutants have strong defects in karyogamy and fail to orient microtubules between parental nuclei in zygotes. In contrast, fus1 mutants show no karyogamy defects. These, and other results suggest that Fus2 defines a novel cell fusion function and subcellular structure that is also required for the alignment of parental nuclei before nuclear fusion.
合子形成是通过紧密协调的细胞和细胞核融合事件发生的。遗传学证据表明,FUS2基因产物在酿酒酵母合子形成过程中促进细胞融合,在细胞壁和质膜融合时或之前与Fus1质膜蛋白协同发挥作用。在此,我们报告了FUS2基因的序列、Fus2蛋白的定位,并表明fus1和fus2突变体在细胞融合方面具有不同的缺陷。FUS2编码一个由617个残基组成的独特开放阅读框,仅在单倍体细胞中响应交配信息素时表达。与在细胞融合中的作用一致,Fus2蛋白定位于离散结构,这些结构可能起源于细胞骨架或囊泡,聚集在信息素诱导的芽体尖端以及合子中配对细胞的交界处。Fus2预计是一种卷曲螺旋蛋白,并与100,000 g的沉淀分离,这表明它与细胞骨架、膜或其他大分子结构相关。Fus2可能与细胞融合过程中细胞核排列所涉及的结构相互作用,因为fus2突变体在核融合方面有严重缺陷,并且无法在合子中亲代细胞核之间定向微管。相比之下,fus1突变体没有核融合缺陷。这些以及其他结果表明,Fus2定义了一种新的细胞融合功能和亚细胞结构,这在核融合之前亲代细胞核的排列中也是必需的。