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在生长中的构巢曲霉菌丝中,高尔基体类似物的快速尖端定向移动表明了一种与生长相关物质传递的机制。

Rapid tip-directed movement of Golgi equivalents in growing Aspergillus nidulans hyphae suggests a mechanism for delivery of growth-related materials.

作者信息

Hubbard Michelle A, Kaminskyj Susan G W

机构信息

Department of Biology, University of Saskatchewan, 112 Science Place, Saskatoon, SK S7N 5E2, Canada.

出版信息

Microbiology (Reading). 2008 May;154(Pt 5):1544-1553. doi: 10.1099/mic.0.2007/014811-0.

Abstract

Golgi equivalents (GEs) process materials in the fungal secretory pathway. Despite the importance of localized secretion in fungal tip growth, GE behaviour in living hyphae has not been documented. The distribution was monitored of an Aspergillus nidulans putative GE-associated protein, CopA, tagged with GFP (CopA-GFP). This co-localized with a Golgi body/GE marker established in other systems, alpha-2,6-sialyltransferase, tagged with red fluorescent protein (ST-RFP). CopA-GFP and ST-RFP distributions responded similarly to brefeldin A, which impairs Golgi/GE trafficking. We used a CopA-GFP, hypA1 strain to study GE distribution and behaviour in growing A. nidulans hyphae. This strain has a wild-type phenotype at 28 degrees C, can be manipulated by changing growth temperature or by use of cytoskeleton inhibitors, and its GE behaviour is consistent with that in a wild-type-morphology strain. A. nidulans GEs were more abundant at hyphal tips than subapically, and showed saltatory motility in all directions. Anterograde GE movements predominated. These were positively correlated with, but at least 10-fold faster than, hyphal growth rate, under all growth and experimental conditions investigated. The actin inhibitor latrunculin B reduced both anterograde GE movement and hyphal growth rate, whereas the microtubule (MT) depolymerizer benomyl increased anterograde GE movement and decreased hyphal growth rate. The MT stabilizer taxol increased A. nidulans GE movement but not hyphal growth rate. A. nidulans GE motility appears to have a complex dependence on both actin and MTs. We present a model for apical delivery of growth materials in which A. nidulans GEs play a role in long-distance transport.

摘要

高尔基体类似物(GEs)在真菌分泌途径中处理物质。尽管局部分泌在真菌顶端生长中很重要,但尚未记录到活菌丝中GE的行为。监测了用绿色荧光蛋白(CopA-GFP)标记的构巢曲霉假定的GE相关蛋白CopA的分布。它与在其他系统中建立的高尔基体/GE标记物α-2,6-唾液酸转移酶共定位,该标记物用红色荧光蛋白(ST-RFP)标记。CopA-GFP和ST-RFP的分布对布雷菲德菌素A的反应相似,布雷菲德菌素A会损害高尔基体/GE的运输。我们使用CopA-GFP、hypA1菌株来研究生长中的构巢曲霉菌丝中GE的分布和行为。该菌株在28摄氏度时具有野生型表型,可以通过改变生长温度或使用细胞骨架抑制剂进行操作,其GE行为与野生型形态菌株一致。构巢曲霉的GE在菌丝顶端比在亚顶端更丰富,并在各个方向上表现出跳跃式运动。顺行性GE运动占主导。在所有研究的生长和实验条件下,这些运动与菌丝生长速率呈正相关,但比菌丝生长速率至少快10倍。肌动蛋白抑制剂拉特罗霉素B降低了顺行性GE运动和菌丝生长速率,而微管(MT)解聚剂苯菌灵增加了顺行性GE运动并降低了菌丝生长速率。MT稳定剂紫杉醇增加了构巢曲霉GE的运动,但没有增加菌丝生长速率。构巢曲霉GE的运动似乎对肌动蛋白和MT都有复杂的依赖性。我们提出了一个生长物质顶端递送的模型,其中构巢曲霉的GE在长距离运输中发挥作用。

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