The COOH-terminal domain of Myo2p, a yeast myosin V, has a direct role in secretory vesicle targeting.

The COOH-terminal domain of Myo2p, a yeast myosin V, has a direct role in secretory vesicle targeting.
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DOI:
10.1083/jcb.147.4.791
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发表时间:
1999-11-15
期刊:
The Journal of cell biology
影响因子:
--
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--
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其他
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MYO 2编码V型肌球蛋白重链,其是将空泡和分泌囊泡靶向酵母生长芽所需的。在这里,我们描述了新的myo2等位基因在COOH末端尾域含有条件性致死突变。在5分钟内转移到限制性温度,极化分布的分泌囊泡被废除,而不影响肌动蛋白或突变体Myo2p的分布,表明尾部有一个直接的作用,在囊泡靶向。我们还表明,肌动蛋白电缆依赖性易位Myo2p的生长位点不需要分泌囊泡货物。虽然含有Myo2p尾的融合蛋白也集中在生长位点,但这种积累取决于分泌囊泡的极化递送,这意味着Myo2p尾与分泌囊泡结合。大多数新的突变改变了与脊椎动物肌球蛋白Vs保守的Myo2p尾区,但与Myo4p不同,Myo4p参与mRNA转运,遗传数据表明,该尾区与Smy1p(驱动蛋白同源物)和Sec4p(囊泡相关Rab蛋白)相互作用。这些数据支持一个模型,其中Myo2p尾栓分泌囊泡,和电机运输他们的极化肌动蛋白电缆的网站胞吐。
MYO2 encodes a type V myosin heavy chain needed for the targeting of vacuoles and secretory vesicles to the growing bud of yeast. Here we describe new myo2 alleles containing conditional lethal mutations in the COOH-terminal tail domain. Within 5 min of shifting to the restrictive temperature, the polarized distribution of secretory vesicles is abolished without affecting the distribution of actin or the mutant Myo2p, showing that the tail has a direct role in vesicle targeting. We also show that the actin cable–dependent translocation of Myo2p to growth sites does not require secretory vesicle cargo. Although a fusion protein containing the Myo2p tail also concentrates at growth sites, this accumulation depends on the polarized delivery of secretory vesicles, implying that the Myo2p tail binds to secretory vesicles. Most of the new mutations alter a region of the Myo2p tail conserved with vertebrate myosin Vs but divergent from Myo4p, the myosin V involved in mRNA transport, and genetic data suggest that the tail interacts with Smy1p, a kinesin homologue, and Sec4p, a vesicle-associated Rab protein. The data support a model in which the Myo2p tail tethers secretory vesicles, and the motor transports them down polarized actin cables to the site of exocytosis.
DOI: 10.1016/s0092-8674(05)80080-7
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