Cytoskeleton-dependent polarized secretion of arylsulfatase in the unicellular green alga, Chlamydomonas reinhardtii
Cytoskeleton-dependent polarized secretion of arylsulfatase in the unicellular green alga, Chlamydomonas reinhardtii
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DOI：
10.1016/j.plantsci.2004.02.003
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发表时间：
2004-06-01
期刊：
影响因子：
5.2
通讯作者：
Kato, S
中科院分区：
文献类型：
--
作者：
Kagiwada, S;Nakamae, I;Kato, S
在绿色衣藻中，蛋白质分泌在细胞壁生物合成、交配过程和应激反应中起着重要作用，但在分子水平上还没有得到很好的研究。在这里，我们研究了衣原体蛋白分泌的细胞骨架的依赖性，通过检查的影响，细胞分裂素破坏试剂。通过芳基硫酸酯酶（ARS）的分泌来监测蛋白质转运活性，ARS是一种糖蛋白，其响应于硫剥夺而合成并转运至周质空间。微管破坏剂，安磺灵和诺考达唑，抑制ARS分泌的剂量依赖性方式，而肌动蛋白破坏剂，细胞松弛素B和D，诱导高达1.4倍的ARS分泌增加。与布雷菲德菌素A（Brefeldin A，BFA）不同的是，oryzatin对细胞内ARS分布的影响是破坏内质网向高尔基体的蛋白质转运。使用抗ARS多克隆抗体的免疫荧光分析揭示了在硫耗尽培养基中培养的细胞的细胞表面的前侧上的特异性点状染色。当用安磺灵处理细胞时，未观察到这种染色。这些结果表明，衣原体蛋白的分泌是依赖于微管和肌动蛋白相关的细胞骨架抑制。这种细胞骨架依赖性类似于哺乳动物细胞中发现的，而不是植物细胞中发现的。(C)2004爱思唯尔爱尔兰有限公司保留所有权利。
In the green alga Chlamydomonas reinhardtii, protein secretion has not been well investigated at the molecular level although the process plays important roles in cell wall biogenesis, the mating process, and stress response. Here, we studied the cytoskeleton dependence of Chlamydomonas protein secretion by examining the effects of cytoskeleton-disrupting reagents. Protein transport activity was monitored by the secretion of arylsulfatase (ARS), a glycoprotein which is synthesized in response to sulfur deprivation and transported to the periplasmic space. Microtubule-disrupting reagents, oryzalin and nocodazole, inhibited ARS secretion in a dose-dependent manner, while actin-disrupting reagents, cytochalasins B and D, induced up to a 1.4-fold increase in ARS secretion. The effect of oryzatin on the intracellular ARS distribution was different from that of brefeldin A (BFA), which disrupts the ER to Golgi protein transport. Immunofluorescence analysis using an anti-ARS polyclonal antibody revealed specific punctate staining on the anterior side of the cell surface of cells cultured in sulfur-depleted medium. This staining was not observed when the cells were treated with oryzalin. These results suggest that Chlamydomonas protein secretion is dependent on microtubules and inhibited by actin-related cytoskeletons. This cytoskeleton dependence is similar to that found in mammalian cells rather than that found in plant cells. (C) 2004 Elsevier Ireland Ltd. All rights reserved.