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Phenotypic and Cell Wall Proteomic Characterization of a DDR48 Mutant Candida albicans Strain

机译:DDR48突变蛋白念珠菌菌株的表型和细胞壁蛋白质重表征

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摘要

Candida albicans is an opportunistic fungus possessing multiple virulence factors controlling pathogenicity. Cell wall proteins are the most important among these factors, being the first elements contacting the host. Ddr48 is a cell wall protein consisting of 212 amino acids. A DDR48 haploinsufficient mutant strain was previously found necessary for proper oxidative stress response and drug resistance. In this study, we aimed to further elucidate the role of Ddr48 by performing additional phenotypic characterization assays. A combinatory proteomic and bioinformatics approach was also undertaken to determine differentially expressed cell wall proteins. Results showed that the mutant strain exhibited a 10% decrease in adhesion mirrored by a 20% decrease in biofilm formation, and slight sensitivity to menadione, diamide, and SDS. Both strains showed similar hyphae formation, virulence, temperature tolerance, and calcofluor white and Congo red sensitivities. Furthermore, a total of 8 and 10 proteins were identified exclusively in the wild-type strain grown under filamentous and non-filamentous conditions respectively. Results included proteins responsible for superoxide stress resistance (Sod4 and Sod6), adhesion (Als3, Hyr4, Pmt1, and Utr2), biofilm formation (Hsp90, Ece1, Rim9, Ipp1, and Pra1) and cell wall integrity (Utr2 and Pga4). The lack of detection of these proteins in the mutant strain correlates with the observed phenotypes.
机译:念珠菌白葡萄酒是一种具有控制致病性的多种毒力因子的机会真菌。细胞壁蛋白是这些因素中最重要的,是联系宿主的第一元素。 DDR48是由212个氨基酸组成的细胞壁蛋白。先前发现了DDR48包发型突变菌株,以获得适当的氧化应激反应和耐药性。在这项研究中,我们旨在通过进行额外的表型表征测定来进一步阐明DDR48的作用。还进行了组合蛋白质组学和生物信息学方法,以确定差异表达的细胞壁蛋白。结果表明,突变菌株在生物膜形成下降20%降低的粘合剂减少10%降低,对男女淋,酰胺和SDS的微小敏感性。两种菌株显示出类似的菌丝形成,毒力,耐腐蚀和钙荧光白色和刚果红色敏感性。此外,分别在丝状和非丝状条件下的野生型菌株中仅鉴定了总共8和10个蛋白质。结果包括负载超氧化物胁迫(SOD4和SOD6)的蛋白质,粘附(ALS3,HYR4,PMT1和UTR2),生物膜形成(HSP90,ECE1,RIM9,IPP1和PRA1)和细胞壁完整性(UTR2和PGA4)。在突变菌株中缺乏这些蛋白质的检测与观察到的表型相关。

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