Castillon Guillaume A, Adames Neil R, Rosello Caroline H, Seidel Hannah S, Longtine Mark S, Cooper John A, Heil-Chapdelaine Richard A
Department of Cell Biology, Washington University, St. Louis, MO 63110, USA.
Curr Biol. 2003 Apr 15;13(8):654-8. doi: 10.1016/s0960-9822(03)00247-1.
In Saccharomyces cerevisiae, the spindle position checkpoint ensures that cells do not exit mitosis until the mitotic spindle moves into the mother/bud neck and thus guarantees that each cell receives one nucleus [1-6]. Mitotic exit is controlled by the small G protein Tem1p. Tem1p and its GTPase activating protein (GAP) Bub2p/Bfa1p are located on the daughter-bound spindle pole body. The GEF Lte1p is located in the bud. This segregation helps keep Tem1p in its inactive GDP state until the spindle enters the neck. However, the checkpoint functions without Lte1p and apparently senses cytoplasmic microtubules in the mother/bud neck [7-9]. To investigate this mechanism, we examined mutants defective for septins, which compose a ring at the neck [10]. We found that the septin mutants sep7Delta and cdc10Delta are defective in the checkpoint. When movement of the spindle into the neck was delayed, mitotic exit occurred, inappropriately leaving both nuclei in the mother. In sep7Delta and cdc10Delta mutants, Lte1p is mislocalized to the mother. In sep7Delta, but not cdc10Delta, mutants, inappropriate mitotic exit depends on Lte1p. These results suggest that septins serve as a diffusion barrier for Lte1p, and that Cdc10p is needed for the septin ring to serve as a scaffold for a putative microtubule sensor.
在酿酒酵母中,纺锤体位置检查点可确保细胞在有丝分裂纺锤体移入母细胞/芽颈之前不会退出有丝分裂,从而保证每个细胞都能获得一个细胞核[1 - 6]。有丝分裂退出由小G蛋白Tem1p控制。Tem1p及其GTP酶激活蛋白(GAP)Bub2p/Bfa1p位于与子细胞相连的纺锤体极体上。鸟嘌呤核苷酸交换因子Lte1p位于芽中。这种分隔有助于使Tem1p保持在无活性的GDP状态,直到纺锤体进入颈部。然而,该检查点在没有Lte1p的情况下也能发挥作用,并且显然能感知母细胞/芽颈中的细胞质微管[7 - 9]。为了研究这一机制,我们检测了隔膜蛋白缺陷的突变体,隔膜蛋白在颈部形成一个环[10]。我们发现隔膜蛋白突变体sep7Δ和cdc10Δ在检查点方面存在缺陷。当纺锤体移入颈部的过程延迟时,有丝分裂退出发生,不恰当地使两个细胞核都留在母细胞中。在sep7Δ和cdc10Δ突变体中,Lte1p定位错误到母细胞中。在sep7Δ突变体中,但不是cdc10Δ突变体中,不适当的有丝分裂退出依赖于Lte1p。这些结果表明,隔膜蛋白作为Lte1p的扩散屏障,并且Cdc10p是隔膜环作为假定微管传感器支架所必需的。