Nowrousian Minou, Frank Sandra, Koers Sandra, Strauch Peter, Weitner Thomas, Ringelberg Carol, Dunlap Jay C, Loros Jennifer J, Kück Ulrich
Lehrstuhl für Allgemeine und Molekulare Botanik, Ruhr-Universität Bochum, Bochum, Germany.
Mol Microbiol. 2007 May;64(4):923-37. doi: 10.1111/j.1365-2958.2007.05694.x.
The filamentous fungus Sordaria macrospora develops complex fruiting bodies (perithecia) to propagate its sexual spores. Here, we present an analysis of the sterile mutant pro41 that is unable to produce mature fruiting bodies. The mutant carries a deletion of 4 kb and is complemented by the pro41 open reading frame that is contained within the region deleted in the mutant. In silico analyses predict PRO41 to be an endoplasmic reticulum (ER) membrane protein, and a PRO41-EGFP fusion protein colocalizes with ER-targeted DsRED. Furthermore, Western blot analysis shows that the PRO41-EGFP fusion protein is present in the membrane fraction. A fusion of the predicted N-terminal signal sequence of PRO41 with EGFP is secreted out of the cell, indicating that the signal sequence is functional. pro41 transcript levels are upregulated during sexual development. This increase in transcript levels was not observed in the sterile mutant pro1 that lacks a transcription factor gene. Moreover, microarray analysis of gene expression in the mutants pro1, pro41 and the pro1/41 double mutant showed that pro41 is partly epistatic to pro1. Taken together, these data show that PRO41 is a novel ER membrane protein essential for fruiting body formation in filamentous fungi.
丝状真菌大孢粪壳菌会形成复杂的子实体(子囊壳)来传播其有性孢子。在此,我们对无法产生成熟子实体的不育突变体pro41进行了分析。该突变体缺失了4 kb的片段,并且由突变体中缺失区域内包含的pro41开放阅读框进行互补。计算机分析预测PRO41是一种内质网(ER)膜蛋白,并且PRO41 - EGFP融合蛋白与内质网靶向的DsRED共定位。此外,蛋白质印迹分析表明PRO41 - EGFP融合蛋白存在于膜组分中。将预测的PRO41的N端信号序列与EGFP融合后可分泌到细胞外,这表明该信号序列具有功能。在有性发育过程中,pro41转录水平上调。在缺乏转录因子基因的不育突变体pro1中未观察到转录水平的这种增加。此外,对突变体pro1、pro41和pro1/41双突变体中的基因表达进行微阵列分析表明,pro41对pro1具有部分上位性。综上所述,这些数据表明PRO41是丝状真菌中形成子实体所必需的一种新型内质网膜蛋白。