So Kum-Kang, Kim Dae-Hyuk
Department of Molecular Biology, Department of Bioactive Material Sciences, Institute for Molecular Biology and Genetics, Chonbuk National University, Jeonju 54896, Korea.
Mycobiology. 2017 Dec;45(4):362-369. doi: 10.5941/MYCO.2017.45.4.362. Epub 2017 Dec 31.
We assessed the regulation of cryparin, a class II hydrophobin, using three representative mitogen-activated protein kinase (MAPK) pathways in . Mutation of the gene, an ortholog of yeast in the cell wall integrity (CWI) pathway, resulted in a dramatic decrease in cryparin production. Similarly, a mutant of the gene, a MAP kinase kinase kinase gene in the CWI pathway, showed decreased cryparin production. Additionally, mutation of the 1 gene, an ortholog of yeast , showed decreased cryparin production. However, mutation of the 2 gene, an ortholog of yeast Kss1/Fus3, showed increased cryparin production. The easy-wet phenotype and accumulation of the cryparin transcript in corresponding mutants were consistent with the cryparin production results. analysis of the promoter region of the cryparin gene revealed the presence of binding motifs related to downstream transcription factors of CWI, HOG1, and pheromone responsive pathways including MADS-box- and Ste12-binding domains. Real-time reverse transcriptase PCR analyses indicated that both , an ortholog of yeast in the CWI pathway, and , an ortholog of yeast in the mating pathway, showed significantly reduced transcription levels in the mutant strains showing lower cryparin production in . However, the transcription of , an ortholog of yeast , did not correlate with that of the mutant strains showing downregulation of cryparin. These results indicate that three representative MAPK pathways played a role in regulating cryparin production. However, regulation varied depending on the MAPK pathways: the CWI and HOG1 pathways were stimulatory, whereas the pheromone-responsive MAPK was repressive.
我们使用三种代表性的丝裂原活化蛋白激酶（MAPK）途径评估了II类疏水蛋白cryparin的调控。细胞壁完整性（CWI）途径中酵母基因的直系同源基因发生突变，导致cryparin产量大幅下降。同样，CWI途径中的MAP激酶激酶激酶基因发生突变，cryparin产量也下降。此外，酵母直系同源基因1发生突变，cryparin产量也下降。然而，酵母Kss1/Fus3直系同源基因2发生突变，cryparin产量增加。相应突变体中cryparin转录本的易湿表型和积累与cryparin产量结果一致。对cryparin基因启动子区域的分析表明，存在与CWI、HOG1下游转录因子以及包括MADS盒和Ste12结合域在内的信息素反应途径相关的结合基序。实时逆转录聚合酶链反应分析表明，CWI途径中酵母直系同源基因和交配途径中酵母直系同源基因在cryparin产量较低的突变菌株中均显示出显著降低的转录水平。然而，酵母直系同源基因的转录与cryparin下调的突变菌株的转录无关。这些结果表明，三种代表性的MAPK途径在调节cryparin产量中发挥了作用。然而，调控因MAPK途径而异：CWI和HOG1途径具有刺激作用，而信息素反应性MAPK具有抑制作用。
