Comparison of Cell Wall Localization among Pir Family Proteins and Functional Dissection of the Region Required for Cell Wall Binding and Bud Scar Recruitment of Pir1p
Comparison of Cell Wall Localization among Pir Family Proteins and Functional Dissection of the Region Required for Cell Wall Binding and Bud Scar Recruitment of Pir1p
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DOI：
10.1128/ec.4.11.1872-1881.2005
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发表时间：
2005-11
期刊：
影响因子：
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通讯作者：
Toru Sumita;T. Yoko-o;Y. Shimma;Y. Jigami
中科院分区：
文献类型：
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作者：
Toru Sumita;T. Yoko-o;Y. Shimma;Y. Jigami
摘要我们研究了Pir蛋白家族（Pir 1至Pir 4）的定位，Pir蛋白家族以未知的方式与细胞壁共价连接。与其他Pir蛋白相比，Pir 1 p与单体红色荧光蛋白的融合蛋白在pir 1 Δ细胞中呈簇状分布，表明Pir 1 p定位于芽痕中。进一步的显微镜分析显示，Pir 1 p在芽痕的几丁质环内表达。逐步删除Pir 1 p重复序列的8个单元揭示了一个单元足以使蛋白质结合芽痕，并且Pir 1 p与细胞壁结合的程度取决于这些重复单元的数量。Pir 1 p的重复序列被Pir 4p的重复序列取代的嵌合Pir 1 p的定位揭示了不同蛋白质区域的功能作用，特别是，重复序列是结合细胞壁所需的，C-末端序列是招募芽痕所需的。这是首次报道芽痕含有Pir 1 p等蛋白质作为内部成分。
ABSTRACT We examined the localization of the Pir protein family (Pir1 to Pir4), which is covalently linked to the cell wall in an unknown manner. In contrast to the other Pir proteins, a fusion of Pir1p and monomeric red fluorescent protein distributed in clusters in pir1Δ cells throughout the period of cultivation, indicating that Pir1p is localized in bud scars. Further microscopic analysis revealed that Pir1p is expressed inside the chitin rings of the bud scars. Stepwise deletion of the eight units of the repetitive sequence of Pir1p revealed that one unit is enough for the protein to bind bud scars and that the extent of binding of Pir1p to the cell wall depends on the number of these repetitive units. The localization of a chimeric Pir1p in which the repetitive sequence of Pir1p was replaced with that of Pir4p revealed the functional role of the different protein regions, specifically, that the repetitive sequence is required for binding to the cell wall and that the C-terminal sequence is needed for recruitment to bud scars. This is the first report that bud scars contain proteins like Pir1p as internal components.