Cprp-An Unusual, Repetitive Protein Which Impacts Pleuromutilin Biosynthesis in the Basidiomycete Clitopilus passeckerianus.
Cprp-An Unusual, Repetitive Protein Which Impacts Pleuromutilin Biosynthesis in the Basidiomycete Clitopilus passeckerianus.
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DOI：
10.3389/ffunb.2021.655323
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发表时间：
2021
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查询EST数据库的Clitopilus passeckerianus确定了一个假定的同源物不寻常的压力反应基因从酵母; ddr 48，被上调截短侧耳素生产条件下。这个基因的沉默，命名为cprp，产生了一个群体的转化体，表现出显着减少截短侧耳素生产。用cprp补充酿酒酵母ddr48突变株（菌株Y16748）的尝试由于缺乏可清楚识别的突变表型而受到阻碍，但有趣的是，ddr48或cprp在S. cerevisiae Y16748导致生长速率的显著和可比较的降低。这一观察结果，结合已知的作用DDR48蛋白从一系列真菌物种在营养饥饿和应激反应，提高了可能性，这个家族的蛋白质在触发寡营养生长中发挥作用。通过在C. Passeckerianus在菌丝隔膜附近显示出清晰的定位，并且在较小程度上，在细胞壁附近，这与DDR48作为各种酵母物种中的细胞壁相关蛋白的鉴定一致。据我们所知，这是第一个研究表明DDR48样蛋白在次级代谢产物的调节中起作用，并代表了第一个来自担子菌的DDR48样蛋白。潜在的同源物可以在大部分的Dikarya中鉴定出来，这表明这种不寻常的蛋白质可能在调节真菌的初级和次级代谢中发挥核心作用。
Interrogation of an EST database for Clitopilus passeckerianus identified a putative homolog to the unusual stress response gene from yeast; ddr48, as being upregulated under pleuromutilin production conditions. Silencing of this gene, named cprp, produced a population of transformants which demonstrated significantly reduced pleuromutilin production. Attempts to complement a Saccharomyces cerevisiae ddr48 mutant strain (strain Y16748) with cprp were hampered by the lack of a clearly identifiable mutant phenotype, but interestingly, overexpression of either ddr48 or cprp in S. cerevisiae Y16748 led to a conspicuous and comparable reduction in growth rate. This observation, combined with the known role of DDR48 proteins from a range of fungal species in nutrient starvation and stress responses, raises the possibility that this family of proteins plays a role in triggering oligotrophic growth. Localization studies via the production of a Cprp:GFP fusion protein in C. passeckerianus showed clear localization adjacent to the hyphal septa and, to a lesser extent, cell walls, which is consistent with the identification of DDR48 as a cell wall-associated protein in various yeast species. To our knowledge this is the first study demonstrating that a DDR48-like protein plays a role in the regulation of a secondary metabolite, and represents the first DDR48-like protein from a basidiomycete. Potential homologs can be identified across much of the Dikarya, suggesting that this unusual protein may play a central role in regulating both primary and secondary metabolism in fungi.