Wolfers Simon, Kamerewerd Jens, Nowrousian Minou, Sigl Claudia, Zadra Ivo, Kürnsteiner Hubert, Kück Ulrich, Bloemendal Sandra
Christian Doppler Laboratory for Fungal Biotechnology, Lehrstuhl für Allgemeine und Molekulare Botanik, Ruhr-Universität Bochum, Bochum, Germany.
J Basic Microbiol. 2015 Apr;55(4):480-9. doi: 10.1002/jobm.201400588. Epub 2015 Jan 2.
The fungal velvet complex is a light-dependent master regulator of secondary metabolism and development in the major penicillin producer, Penicillium chrysogenum. However, the light-dependent mechanism is unclear. To identify velvet-dependent transcriptional regulators that show light-regulated expression, we performed microarray hybridizations with RNA isolated from P. chrysogenum ΔPcku70 cultures grown under 13 different long-term, light-dependent growth conditions. We compared these expression data to data from two velvet complex deletion mutants; one lacked a subunit of the velvet complex (ΔPcvelA), and the other lacked a velvet-associated protein (ΔPclaeA). We sought to identify genes that were up-regulated in light, but down-regulated in ΔPcvelA and ΔPclaeA. We identified 148 co-regulated genes that displayed this regulatory pattern. In silico analyses of the co-regulated genes identified six proteins with fungal-specific transcription factor domains. Among these, we selected the bZIP transcription factor, PcAtfA, for functional characterization in deletion and complementation strains. Our data clearly indicates that PcAtfA governs spore germination. This comparative analysis of different microarray hybridization data sets provided results that may be useful for identifying genes for future functional analyses.
真菌绒毛复合体是主要青霉素生产菌产黄青霉中次级代谢和发育的光依赖性主调控因子。然而,其光依赖性机制尚不清楚。为了鉴定显示光调控表达的绒毛依赖性转录调控因子,我们用从产黄青霉ΔPcku70培养物中分离的RNA进行了微阵列杂交,该培养物在13种不同的长期光依赖性生长条件下生长。我们将这些表达数据与来自两个绒毛复合体缺失突变体的数据进行了比较;一个缺失绒毛复合体的一个亚基(ΔPcvelA),另一个缺失与绒毛相关的蛋白(ΔPclaeA)。我们试图鉴定在光照下上调,但在ΔPcvelA和ΔPclaeA中下调的基因。我们鉴定出148个显示这种调控模式的共调控基因。对共调控基因的计算机分析鉴定出六种具有真菌特异性转录因子结构域的蛋白质。其中,我们选择了bZIP转录因子PcAtfA,在缺失和互补菌株中进行功能表征。我们的数据清楚地表明,PcAtfA控制孢子萌发。这种对不同微阵列杂交数据集的比较分析提供了可能有助于鉴定未来功能分析基因的结果。