Guerriero Gea, Silvestrini Lucia, Obersriebnig Michael, Hausman Jean-Francois, Strauss Joseph, Ezcurra Inés
Department of Applied Genetics and Cell Biology, Fungal Genetics and Genomics Unit, University of Natural Resources and Life Sciences Vienna (BOKU), Bioresources and Technologies Campus Tulln-Technopol, Tulln/Donau A-3430, Austria.
Environmental Research and Innovation (ERIN), Luxembourg Institute of Science and Technology (LIST), Esch/Alzette L-4362, Luxembourg.
Int J Mol Sci. 2016 Jun 29;17(7):1031. doi: 10.3390/ijms17071031.
WD40 repeat (WDR) proteins are pleiotropic molecular hubs. We identify a WDR gene that is a conserved genomic neighbor of a chitin synthase gene in Ascomycetes. The WDR gene is unique to fungi and plants, and was called Fungal Plant WD (FPWD). FPWD is within a cell wall metabolism gene cluster in the Ascomycetes (Pezizomycotina) comprising chsD, a Chs activator and a GH17 glucanase. The FPWD, AN1556.2 locus was deleted in Aspergillus nidulans strain SAA.111 by gene replacement and only heterokaryon transformants were obtained. The re-annotation of Aspergilli genomes shows that AN1556.2 consists of two tightly linked separate genes, i.e., the WDR gene and a putative beta-flanking gene of unknown function. The WDR and the beta-flanking genes are conserved genomic neighbors localized within a recently identified metabolic cell wall gene cluster in genomes of Aspergilli. The heterokaryons displayed increased susceptibility to drugs affecting the cell wall, and their phenotypes, observed by optical, confocal, scanning electron and atomic force microscopy, suggest cell wall alterations. Quantitative real-time PCR shows altered expression of some cell wall-related genes. The possible implications on cell wall biosynthesis are discussed.
WD40重复(WDR)蛋白是多效性分子枢纽。我们鉴定出一个WDR基因,它是子囊菌中几丁质合酶基因的保守基因组邻接基因。该WDR基因是真菌和植物所特有的,被称为真菌植物WD(FPWD)。FPWD位于子囊菌(粪壳菌纲)的一个细胞壁代谢基因簇内,该基因簇包含chsD、一种几丁质合酶激活剂和一种GH17葡聚糖酶。通过基因替换在构巢曲霉SAA.111菌株中缺失了FPWD(AN1556.2位点),并且只获得了异核体转化体。对曲霉基因组的重新注释表明,AN1556.2由两个紧密连锁的独立基因组成,即WDR基因和一个功能未知的假定β侧翼基因。WDR基因和β侧翼基因是保守的基因组邻接基因,定位于曲霉基因组中最近鉴定出の代谢细胞壁基因簇内。异核体对影响细胞壁的药物表现出更高的敏感性,通过光学显微镜、共聚焦显微镜、扫描电子显微镜和原子力显微镜观察到的它们的表型表明细胞壁发生了改变。定量实时PCR显示一些细胞壁相关基因的表达发生了变化。文中讨论了其对细胞壁生物合成可能产生的影响。