白色念珠菌胞囊Sec15亚基的功能分析。

Eukaryotic Cell Pub Date : 2015-12-01 Epub Date: 2015-10-09 DOI:10.1128/EC.00147-15
Alba A Chavez-Dozal, Stella M Bernardo, Hallie S Rane, Samuel A Lee
{"title":"白色念珠菌胞囊Sec15亚基的功能分析。","authors":"Alba A Chavez-Dozal,&nbsp;Stella M Bernardo,&nbsp;Hallie S Rane,&nbsp;Samuel A Lee","doi":"10.1128/EC.00147-15","DOIUrl":null,"url":null,"abstract":"<p><p>In prior studies of exocyst-mediated late secretion in Candida albicans, we have determined that Sec6 contributes to cell wall integrity, secretion, and filamentation. A conditional mutant lacking SEC6 expression exhibits markedly reduced lateral hyphal branching. In addition, lack of the related t-SNAREs Sso2 and Sec9 also leads to defects in secretion and filamentation. To further understand the role of the exocyst in the fundamental processes of polarized secretion and filamentation in C. albicans, we studied the exocyst subunit Sec15. Since Saccharomyces cerevisiae SEC15 is essential for viability, we generated a C. albicans conditional mutant strain in which SEC15 was placed under the control of a tetracycline-regulated promoter. In the repressed state, cell death occurred after 5 h in the tetR-SEC15 strain. Prior to this time point, the tetR-SEC15 mutant was markedly defective in Sap and lipase secretion and demonstrated increased sensitivity to Zymolyase and chitinase. Notably, tetR-SEC15 mutant hyphae were characterized by a hyperbranching phenotype, in direct contrast to strain tetR-SEC6, which had minimal lateral branching. We further studied the localization of the Spitzenkörper, polarisomes, and exocysts in the tetR-SEC15 and tetR-SEC6 mutants during filamentation. Mlc1-GFP (marking the Spitzenkörper), Spa2-GFP (the polarisome), and Exo70-GFP (exocyst) localizations were normal in the tetR-SEC6 mutant, whereas these structures were mislocalized in the tetR-SEC15 mutant. Following alleviation of gene repression by removing doxycycline, first Spitzenkörper, then polarisome, and finally exocyst localizations were recovered sequentially. These results indicate that the exocyst subunits Sec15 and Sec6 have distinct roles in mediating polarized secretion and filamentation in C. albicans. </p>","PeriodicalId":11891,"journal":{"name":"Eukaryotic Cell","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2015-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1128/EC.00147-15","citationCount":"7","resultStr":"{\"title\":\"Functional Analysis of the Exocyst Subunit Sec15 in Candida albicans.\",\"authors\":\"Alba A Chavez-Dozal,&nbsp;Stella M Bernardo,&nbsp;Hallie S Rane,&nbsp;Samuel A Lee\",\"doi\":\"10.1128/EC.00147-15\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>In prior studies of exocyst-mediated late secretion in Candida albicans, we have determined that Sec6 contributes to cell wall integrity, secretion, and filamentation. A conditional mutant lacking SEC6 expression exhibits markedly reduced lateral hyphal branching. In addition, lack of the related t-SNAREs Sso2 and Sec9 also leads to defects in secretion and filamentation. To further understand the role of the exocyst in the fundamental processes of polarized secretion and filamentation in C. albicans, we studied the exocyst subunit Sec15. Since Saccharomyces cerevisiae SEC15 is essential for viability, we generated a C. albicans conditional mutant strain in which SEC15 was placed under the control of a tetracycline-regulated promoter. In the repressed state, cell death occurred after 5 h in the tetR-SEC15 strain. Prior to this time point, the tetR-SEC15 mutant was markedly defective in Sap and lipase secretion and demonstrated increased sensitivity to Zymolyase and chitinase. Notably, tetR-SEC15 mutant hyphae were characterized by a hyperbranching phenotype, in direct contrast to strain tetR-SEC6, which had minimal lateral branching. We further studied the localization of the Spitzenkörper, polarisomes, and exocysts in the tetR-SEC15 and tetR-SEC6 mutants during filamentation. Mlc1-GFP (marking the Spitzenkörper), Spa2-GFP (the polarisome), and Exo70-GFP (exocyst) localizations were normal in the tetR-SEC6 mutant, whereas these structures were mislocalized in the tetR-SEC15 mutant. Following alleviation of gene repression by removing doxycycline, first Spitzenkörper, then polarisome, and finally exocyst localizations were recovered sequentially. These results indicate that the exocyst subunits Sec15 and Sec6 have distinct roles in mediating polarized secretion and filamentation in C. albicans. </p>\",\"PeriodicalId\":11891,\"journal\":{\"name\":\"Eukaryotic Cell\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2015-12-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1128/EC.00147-15\",\"citationCount\":\"7\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Eukaryotic Cell\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1128/EC.00147-15\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2015/10/9 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Eukaryotic Cell","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1128/EC.00147-15","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2015/10/9 0:00:00","PubModel":"Epub","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 7

摘要

在之前关于白色念珠菌胞囊介导的晚期分泌的研究中,我们已经确定Sec6有助于细胞壁的完整性、分泌和丝化。缺乏SEC6表达的条件突变体表现出明显减少的菌丝分枝。此外,缺乏相关的t-SNAREs Sso2和Sec9也会导致分泌和成丝缺陷。为了进一步了解胞囊在白色念珠菌极化分泌和丝化的基本过程中的作用,我们研究了胞囊亚基Sec15。由于酿酒酵母(Saccharomyces cerevisiae)的SEC15对其生存能力至关重要,我们产生了一个白色念珠菌条件突变株,其中SEC15置于四环素调节启动子的控制下。在抑制状态下,细胞在5 h后死亡。在此时间点之前,ter - sec15突变体在汁液和脂肪酶分泌方面明显缺陷,对酶解酶和几丁质酶的敏感性增加。值得注意的是,ter - sec15突变体菌丝具有超分支表型,与菌株ter - sec6形成鲜明对比,后者具有最小的侧分支。我们进一步研究了tr - sec15和tr - sec6突变体在成丝过程中Spitzenkörper、极化体和胞囊的定位。Mlc1-GFP(标记Spitzenkörper), Spa2-GFP(极化体)和Exo70-GFP(外囊)定位在ter - sec6突变体中是正常的,而这些结构在ter - sec15突变体中定位错误。通过去除强力霉素减轻基因抑制后,首先Spitzenkörper,然后极化体,最后囊胞定位依次恢复。这些结果表明,胞囊亚基Sec15和Sec6在介导白色念珠菌极化分泌和丝化过程中具有不同的作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Functional Analysis of the Exocyst Subunit Sec15 in Candida albicans.

Functional Analysis of the Exocyst Subunit Sec15 in Candida albicans.

Functional Analysis of the Exocyst Subunit Sec15 in Candida albicans.

Functional Analysis of the Exocyst Subunit Sec15 in Candida albicans.

In prior studies of exocyst-mediated late secretion in Candida albicans, we have determined that Sec6 contributes to cell wall integrity, secretion, and filamentation. A conditional mutant lacking SEC6 expression exhibits markedly reduced lateral hyphal branching. In addition, lack of the related t-SNAREs Sso2 and Sec9 also leads to defects in secretion and filamentation. To further understand the role of the exocyst in the fundamental processes of polarized secretion and filamentation in C. albicans, we studied the exocyst subunit Sec15. Since Saccharomyces cerevisiae SEC15 is essential for viability, we generated a C. albicans conditional mutant strain in which SEC15 was placed under the control of a tetracycline-regulated promoter. In the repressed state, cell death occurred after 5 h in the tetR-SEC15 strain. Prior to this time point, the tetR-SEC15 mutant was markedly defective in Sap and lipase secretion and demonstrated increased sensitivity to Zymolyase and chitinase. Notably, tetR-SEC15 mutant hyphae were characterized by a hyperbranching phenotype, in direct contrast to strain tetR-SEC6, which had minimal lateral branching. We further studied the localization of the Spitzenkörper, polarisomes, and exocysts in the tetR-SEC15 and tetR-SEC6 mutants during filamentation. Mlc1-GFP (marking the Spitzenkörper), Spa2-GFP (the polarisome), and Exo70-GFP (exocyst) localizations were normal in the tetR-SEC6 mutant, whereas these structures were mislocalized in the tetR-SEC15 mutant. Following alleviation of gene repression by removing doxycycline, first Spitzenkörper, then polarisome, and finally exocyst localizations were recovered sequentially. These results indicate that the exocyst subunits Sec15 and Sec6 have distinct roles in mediating polarized secretion and filamentation in C. albicans.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Eukaryotic Cell
Eukaryotic Cell 生物-微生物学
自引率
0.00%
发文量
0
审稿时长
1 months
期刊介绍: Eukaryotic Cell (EC) focuses on eukaryotic microbiology and presents reports of basic research on simple eukaryotic microorganisms, such as yeasts, fungi, algae, protozoa, and social amoebae. The journal also covers viruses of these organisms and their organelles and their interactions with other living systems, where the focus is on the eukaryotic cell. Topics include: - Basic biology - Molecular and cellular biology - Mechanisms, and control, of developmental pathways - Structure and form inherent in basic biological processes - Cellular architecture - Metabolic physiology - Comparative genomics, biochemistry, and evolution - Population dynamics - Ecology
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信