Three DUF1996 Proteins Localize in Vacuoles and Function in Fungal Responses to Multiple Stresses and Metal Ions.
Three DUF1996 Proteins Localize in Vacuoles and Function in Fungal Responses to Multiple Stresses and Metal Ions.
复制标题
三种 DUF1996 蛋白定位于液泡并在真菌对多种应力和金属离子的反应中发挥作用
DOI：
10.1038/srep20566
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发表时间：
2016-02-03
影响因子：
4.6
通讯作者：
Feng MG
中科院分区：
文献类型：
--
作者：
Tong SM;Chen Y;Ying SH;Feng MG
许多带注释的真菌基因组含有高比例的假设蛋白质，带有或不带有未知功能域 (DUF)。在这里，三种新蛋白（342−497 个氨基酸），每个仅包含一个大的 DUF1996（231−250 个残基）区域，具有高度保守的头部 (DPIXXP) 和尾部 (HXDXXXGW) 特征，被表达为 eGFP 标记的融合蛋白，并显示出特异性定位于白僵菌（一种丝状真菌昆虫病原体）的液泡中；因此，这些蛋白质被命名为液泡定位蛋白（VLP）。 VLP在其他昆虫病原性或非昆虫病原性丝状真菌中具有1至3个同源物，但在酵母中没有同源物。大的DUF1996区域可以表示为D-X4-P-X5–6-H-X-H-X3-G-X25–26-D-X-S-X-YW-X-P-X123–203-CP-X39–48-H-X-D-X3-GW；相同的残基可能参与细胞内稳态的质子反向转运系统。三个 VLP 编码基因 (vlp1-3) 的单一缺失增加了真菌对细胞壁扰动、高渗透压、氧化和多种金属离子的敏感性。两个 Δvlp 突变体的分生孢子耐热性下降了约 11%，三个 Δvlp 突变体的 UV-B 抗性下降了 41−57%。所有的变化都通过靶向基因互补得以恢复。然而，缺失并不影响真菌生长、分生孢子形成、毒力或 Cu2+ 敏感性。我们的研究结果揭示了三种白僵菌 VLP 的 DUF1996 区域在多种应激反应和环境适应调节中的作用。
Many annotated fungal genomes harbour high proportions of hypothetical proteins with or without domains of unknown function (DUF). Here, three novel proteins (342−497 amino acids), each containing only a single large DUF1996 (231−250 residues) region with highly conserved head (DPIXXP) and tail (HXDXXXGW) signatures, were expressed as eGFP-tagged fusion proteins and shown to specifically localize in the vacuoles of Beauveria bassiana, a filamentous fungal entomopathogen; therefore, these proteins were named vacuole-localized proteins (VLPs). The VLPs have one to three homologues in other entomopathogenic or non-entomopathogenic filamentous fungi but no homologues in yeasts. The large DUF1996 regions can be formulated as D-X4-P-X5–6-H-X-H-X3-G-X25–26-D-X-S-X-YW-X-P-X123–203-CP-X39–48-H-X-D-X3-GW; the identical residues likely involve in a proton antiport system for intracellular homeostasis. Single deletions of three VLP-coding genes (vlp1–3) increased fungal sensitivities to cell wall perturbation, high osmolarity, oxidation, and several metal ions. Conidial thermotolerance decreased by ~11% in two Δvlp mutants, and UV-B resistance decreased by 41−57% in three Δvlp mutants. All the changes were restored by targeted gene complementation. However, the deletions did not influence fungal growth, conidiation, virulence or Cu2+ sensitivity. Our findings unveiled a role for the DUF1996 regions of three B. bassiana VLPs in the regulation of multiple stress responses and environmental adaptation.