内质网葡萄糖苷酶II编码基因ROT2的沉默对细胞壁和申克孢子菌与宿主相互作用的影响

Luz A López-Ramírez, Iván Martínez-Duncker, Anayeli Márquez-Márquez, Ana P Vargas-Macías, Héctor M Mora-Montes
{"title":"内质网葡萄糖苷酶II编码基因ROT2的沉默对细胞壁和申克孢子菌与宿主相互作用的影响","authors":"Luz A López-Ramírez,&nbsp;Iván Martínez-Duncker,&nbsp;Anayeli Márquez-Márquez,&nbsp;Ana P Vargas-Macías,&nbsp;Héctor M Mora-Montes","doi":"10.3390/jof8111220","DOIUrl":null,"url":null,"abstract":"<p><p><i>Sporothrix schenckii</i> is a member of the <i>Sporothrix</i> pathogenic clade and one of the most common etiological agents of sporotrichosis, a subcutaneous fungal infection that affects both animal and human beings. Like other fungal pathogens, the <i>Sporothrix</i> cell wall is composed of structural polysaccharides and glycoproteins that are covalently modified with both <i>N</i>-linked and <i>O</i>-linked glycans. Thus far, little is known about the <i>N</i>-linked glycosylation pathway in this organism or its contribution to cell wall composition and interaction with the host. Here, we silenced <i>ROT2</i>, which encodes the catalytic subunit of the endoplasmic reticulum α-glucosidase II, a processing enzyme key for the <i>N</i>-linked glycan core processing. Silencing of <i>ROT2</i> led to the accumulation of the Glc<sub>2</sub>Man<sub>9</sub>GlcNAC<sub>2</sub> glycan core at the cell wall and a reduction in the total content of <i>N</i>-linked glycans found in the wall. However, the highly silenced mutants showed a compensatory mechanism with increased content of cell wall <i>O</i>-linked glycans. The phenotype of mutants with intermediate levels of <i>ROT2</i> silencing was more informative, as they showed changes in the cell wall composition and exposure of β-1.3-glucans and chitin at the cell surface. Furthermore, the ability to stimulate cytokine production by human mononuclear cells was affected, along with the phagocytosis by human monocyte-derived macrophages, in a mannose receptor-, complement receptor 3-, and TLR4-dependent stimulation. In an insect model of experimental sporotrichosis, these mutant cells showed virulence attenuation. In conclusion, <i>S. schenckii ROT2</i> is required for proper <i>N</i>-linked glycosylation, cell wall organization and composition, and interaction with the host.</p>","PeriodicalId":520671,"journal":{"name":"Journal of fungi (Basel, Switzerland)","volume":" ","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2022-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9692468/pdf/","citationCount":"2","resultStr":"{\"title\":\"Silencing of <i>ROT2</i>, the Encoding Gene of the Endoplasmic Reticulum Glucosidase II, Affects the Cell Wall and the <i>Sporothrix schenckii</i>-Host Interaction.\",\"authors\":\"Luz A López-Ramírez,&nbsp;Iván Martínez-Duncker,&nbsp;Anayeli Márquez-Márquez,&nbsp;Ana P Vargas-Macías,&nbsp;Héctor M Mora-Montes\",\"doi\":\"10.3390/jof8111220\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><i>Sporothrix schenckii</i> is a member of the <i>Sporothrix</i> pathogenic clade and one of the most common etiological agents of sporotrichosis, a subcutaneous fungal infection that affects both animal and human beings. Like other fungal pathogens, the <i>Sporothrix</i> cell wall is composed of structural polysaccharides and glycoproteins that are covalently modified with both <i>N</i>-linked and <i>O</i>-linked glycans. Thus far, little is known about the <i>N</i>-linked glycosylation pathway in this organism or its contribution to cell wall composition and interaction with the host. Here, we silenced <i>ROT2</i>, which encodes the catalytic subunit of the endoplasmic reticulum α-glucosidase II, a processing enzyme key for the <i>N</i>-linked glycan core processing. Silencing of <i>ROT2</i> led to the accumulation of the Glc<sub>2</sub>Man<sub>9</sub>GlcNAC<sub>2</sub> glycan core at the cell wall and a reduction in the total content of <i>N</i>-linked glycans found in the wall. However, the highly silenced mutants showed a compensatory mechanism with increased content of cell wall <i>O</i>-linked glycans. The phenotype of mutants with intermediate levels of <i>ROT2</i> silencing was more informative, as they showed changes in the cell wall composition and exposure of β-1.3-glucans and chitin at the cell surface. Furthermore, the ability to stimulate cytokine production by human mononuclear cells was affected, along with the phagocytosis by human monocyte-derived macrophages, in a mannose receptor-, complement receptor 3-, and TLR4-dependent stimulation. In an insect model of experimental sporotrichosis, these mutant cells showed virulence attenuation. In conclusion, <i>S. schenckii ROT2</i> is required for proper <i>N</i>-linked glycosylation, cell wall organization and composition, and interaction with the host.</p>\",\"PeriodicalId\":520671,\"journal\":{\"name\":\"Journal of fungi (Basel, Switzerland)\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2022-11-18\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9692468/pdf/\",\"citationCount\":\"2\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of fungi (Basel, Switzerland)\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.3390/jof8111220\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of fungi (Basel, Switzerland)","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.3390/jof8111220","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 2

摘要

申克孢子菌是孢子菌致病性分支的一员,也是孢子菌病最常见的病原之一,孢子菌病是一种影响动物和人类的皮下真菌感染。像其他真菌病原体一样,孢子丝菌细胞壁由结构多糖和糖蛋白组成,它们被n -链和o -链聚糖共价修饰。到目前为止,人们对这种生物中的n -链糖基化途径或其对细胞壁组成和与宿主相互作用的贡献知之甚少。在这里,我们沉默了编码内质网α-葡萄糖苷酶II催化亚基的ROT2, α-葡萄糖苷酶II是n -链聚糖核心加工的关键酶。ROT2的沉默导致Glc2Man9GlcNAC2聚糖核心在细胞壁处的积累,并减少细胞壁中发现的n -链聚糖的总含量。然而,高度沉默的突变体表现出一种代偿机制,即细胞壁o链聚糖含量增加。具有中等水平ROT2沉默的突变体的表型更具信息量,因为它们显示细胞壁组成的变化以及细胞表面β-1.3葡聚糖和几丁质的暴露。此外,在甘露糖受体-、补体受体3-和tlr4依赖性刺激下,人类单核细胞刺激细胞因子产生的能力以及人类单核细胞来源的巨噬细胞的吞噬能力也受到了影响。在实验性孢子菌病的昆虫模型中,这些突变细胞表现出毒力衰减。综上所述,申氏梭菌ROT2是正常的n -链糖基化、细胞壁组织和组成以及与宿主相互作用所必需的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Silencing of <i>ROT2</i>, the Encoding Gene of the Endoplasmic Reticulum Glucosidase II, Affects the Cell Wall and the <i>Sporothrix schenckii</i>-Host Interaction.

Silencing of <i>ROT2</i>, the Encoding Gene of the Endoplasmic Reticulum Glucosidase II, Affects the Cell Wall and the <i>Sporothrix schenckii</i>-Host Interaction.

Silencing of <i>ROT2</i>, the Encoding Gene of the Endoplasmic Reticulum Glucosidase II, Affects the Cell Wall and the <i>Sporothrix schenckii</i>-Host Interaction.

Silencing of ROT2, the Encoding Gene of the Endoplasmic Reticulum Glucosidase II, Affects the Cell Wall and the Sporothrix schenckii-Host Interaction.

Sporothrix schenckii is a member of the Sporothrix pathogenic clade and one of the most common etiological agents of sporotrichosis, a subcutaneous fungal infection that affects both animal and human beings. Like other fungal pathogens, the Sporothrix cell wall is composed of structural polysaccharides and glycoproteins that are covalently modified with both N-linked and O-linked glycans. Thus far, little is known about the N-linked glycosylation pathway in this organism or its contribution to cell wall composition and interaction with the host. Here, we silenced ROT2, which encodes the catalytic subunit of the endoplasmic reticulum α-glucosidase II, a processing enzyme key for the N-linked glycan core processing. Silencing of ROT2 led to the accumulation of the Glc2Man9GlcNAC2 glycan core at the cell wall and a reduction in the total content of N-linked glycans found in the wall. However, the highly silenced mutants showed a compensatory mechanism with increased content of cell wall O-linked glycans. The phenotype of mutants with intermediate levels of ROT2 silencing was more informative, as they showed changes in the cell wall composition and exposure of β-1.3-glucans and chitin at the cell surface. Furthermore, the ability to stimulate cytokine production by human mononuclear cells was affected, along with the phagocytosis by human monocyte-derived macrophages, in a mannose receptor-, complement receptor 3-, and TLR4-dependent stimulation. In an insect model of experimental sporotrichosis, these mutant cells showed virulence attenuation. In conclusion, S. schenckii ROT2 is required for proper N-linked glycosylation, cell wall organization and composition, and interaction with the host.

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
自引率
0.00%
发文量
0
×
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:604180095
Book学术官方微信