The two transmembrane regions of Candida albicans Dfi1 contribute to its biogenesis.
The two transmembrane regions of Candida albicans Dfi1 contribute to its biogenesis.
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DOI：
10.1016/j.bbrc.2017.04.158
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发表时间：
2017-06-17
影响因子：
3.1
通讯作者：
Kumamoto CA
中科院分区：
文献类型：
--
作者：
Herwald SE;Zucchi PC;Tan S;Kumamoto CA
机会致病菌白色念珠菌形成侵入性丝状体，在疾病期间生长到宿主组织中。糖基化的完整质膜蛋白Dfi 1在实验室模型中对侵袭性抑制和小鼠播散性念珠菌病的致死性都很重要。然而，Dfi 1拓扑结构和Dfi 1生物发生的必要结构域是不确定的。序列分析预测Dfi 1含有两个跨膜区，位于N-和C-末端附近。在这次交流中，我们表明，Dfi 1仍然是一个完整的膜蛋白，尽管删除任何预测的跨膜区，而删除这两个区域的结果在一个可溶性蛋白。此外，Dfi 1是适当的定向在膜上，如N-连接的糖基化，观察时，任何跨膜区域被删除，但不存在时，两个跨膜区域被删除。有趣的是，N-末端跨膜区的缺失导致产生两种形式的Dfi 1。大多数蛋白质分子获得了正常的N-连接糖基化，少数群体未能正常N-连接糖基化。通过用一种合成的跨膜序列而不是另一种序列替换N-末端疏水序列来逆转该缺陷。最后，显微镜研究表明，Dfi 1缺乏N-末端跨膜区观察到在细胞周边，全长Dfi 1通常定位，而双截短突变体是弥漫性细胞内。因此，成熟的Dfi 1蛋白含有两个跨膜结构域，这有助于其生物合成。
The opportunistic pathogen Candida albicans forms invasive filaments that grow into host tissues during disease. The glycosylated, integral plasma membrane protein Dfi1 is important for invasive filamentation in a laboratory model, and for lethality in murine disseminated candidiasis. However, Dfi1 topology and essential domains for Dfi1 biogenesis were undefined. Sequence analysis predicted that Dfi1 contains two transmembrane regions, located near the N- and C-termini. In this communication, we show that Dfi1 remains an integral membrane protein despite deletion of either predicted transmembrane region, whereas deletion of both regions results in a soluble protein. Additionally, Dfi1 that was properly oriented in the membrane, as indicated by N-linked glycosylation, was observed when either transmembrane region was deleted, but was absent when both transmembrane regions were deleted. Interestingly, deletion of the N-terminal transmembrane region resulted in production of two forms of Dfi1. Most of the protein molecules acquired normal N-linked glycosylation and a smaller population failed to become normally N-linked glycosylated. This defect was reversed by replacement of the N-terminal hydrophobic sequence with one synthetic transmembrane sequence but not another. Finally, microscopy studies revealed that Dfi1 lacking the N-terminal transmembrane region was observed at the cell periphery, where full-length Dfi1 normally localizes, whereas the double-truncation mutant was diffusely intracellular. Therefore, mature Dfi1 protein contains two transmembrane domains which contribute to its biogenesis.
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DOI：
10.1111/j.1432-1033.1991.tb16166.x
发表时间：
1991-08-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
TRETTER, V;ALTMANN, F;MARZ, L
通讯作者：
MARZ, L
影响因子：
3.7
作者：
Davis, Talya R.;Zucchi, Paola C.;Kumamoto, Carol A.
通讯作者：
Kumamoto, Carol A.
影响因子：
64.8
作者：
MAEDA, T;WURGLERMURPHY, SM;SAITO, H
通讯作者：
SAITO, H
影响因子：
3.4
作者：
Martinez-Duncker, Ivan;Diaz-Jimenez, Diana F;Mora-Montes, Hector M
通讯作者：
Mora-Montes, Hector M
影响因子：
4.8
作者：
Narang, Sandhya S.;Malone, Cheryl L.;Fassler, Jan S.
通讯作者：
Fassler, Jan S.