Min Kyunghun, Son Hokyoung, Lim Jae Yun, Choi Gyung Ja, Kim Jin-Cheol, Harris Steven D, Lee Yin-Won
Department of Agricultural Biotechnology and Center for Fungal Pathogenesis, Seoul National University, Seoul, Republic of Korea.
Eukaryot Cell. 2014 Mar;13(3):427-36. doi: 10.1128/EC.00293-13. Epub 2014 Jan 24.
The survival of cellular organisms depends on the faithful replication and transmission of DNA. Regulatory factor X (RFX) transcription factors are well conserved in animals and fungi, but their functions are diverse, ranging from the DNA damage response to ciliary gene regulation. We investigated the role of the sole RFX transcription factor, RFX1, in the plant-pathogenic fungus Fusarium graminearum. Deletion of rfx1 resulted in multiple defects in hyphal growth, conidiation, virulence, and sexual development. Deletion mutants of rfx1 were more sensitive to various types of DNA damage than the wild-type strain. Septum formation was inhibited and micronuclei were produced in the rfx1 deletion mutants. The results of the neutral comet assay demonstrated that disruption of rfx1 function caused spontaneous DNA double-strand breaks (DSBs). The transcript levels of genes involved in DNA DSB repair were upregulated in the rfx1 deletion mutants. DNA DSBs produced micronuclei and delayed septum formation in F. graminearum. Green fluorescent protein (GFP)-tagged RFX1 localized in nuclei and exhibited high expression levels in growing hyphae and conidiophores, where nuclear division was actively occurring. RNA-sequencing-based transcriptomic analysis revealed that RFX1 suppressed the expression of many genes, including those required for the repair of DNA damage. Taken together, these findings indicate that the transcriptional repressor rfx1 performs crucial roles during normal cell growth by maintaining genome integrity.
细胞生物的存活依赖于DNA的忠实复制和传递。调控因子X(RFX)转录因子在动物和真菌中高度保守,但其功能多样,涵盖从DNA损伤反应到纤毛基因调控等方面。我们研究了植物致病真菌禾谷镰刀菌中唯一的RFX转录因子RFX1的作用。rfx1的缺失导致菌丝生长、分生孢子形成、毒力和有性发育出现多种缺陷。rfx1的缺失突变体比野生型菌株对各种类型的DNA损伤更敏感。rfx1缺失突变体中隔膜形成受到抑制并产生微核。中性彗星试验结果表明,rfx1功能的破坏导致自发的DNA双链断裂(DSB)。参与DNA DSB修复的基因转录水平在rfx1缺失突变体中上调。DNA DSB在禾谷镰刀菌中产生微核并延迟隔膜形成。绿色荧光蛋白(GFP)标记的RFX1定位于细胞核,并在正在进行核分裂的生长菌丝和分生孢子梗中表现出高表达水平。基于RNA测序的转录组分析表明,RFX1抑制了许多基因的表达,包括那些DNA损伤修复所需的基因。综上所述,这些发现表明转录抑制因子rfx1通过维持基因组完整性在正常细胞生长过程中发挥关键作用。