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Transcription factor RFX1 is critical for maintenance of chromosome integrity in Fusarium graminearum : Fusarium graminearum의 전사조절인자 RFX1의 기능연구
DC Field | Value | Language |
---|---|---|
dc.contributor.advisor | 이인원 | - |
dc.contributor.author | 민경훈 | - |
dc.date.accessioned | 2017-07-13T08:21:33Z | - |
dc.date.available | 2017-07-13T08:21:33Z | - |
dc.date.issued | 2014-08 | - |
dc.identifier.other | 000000021923 | - |
dc.identifier.uri | https://hdl.handle.net/10371/119475 | - |
dc.description | 학위논문 (박사)-- 서울대학교 대학원 : 농생명공학부, 2014. 8. 이인원. | - |
dc.description.abstract | 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 F. 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). Transcript levels of genes involved in DNA DSB repair were up-regulated in the rfx1 deletion mutants. DNA DSBs produced micronuclei and delayed septum formation in F. graminearum. 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. | - |
dc.description.tableofcontents | ABSTRACT 1
INTRODUCTION 7 MATERIALS AND METHODS 11 I. Strains and culture conditions 11 II. Nucleic manipulation, primers, and PCR conditions 11 III. Sexual crosses 16 IV. Fungal transformation 16 V. Fluorescence microscopy 17 VI. Virulence assays 17 VII. Toxin analysis 18 VIII. Neutral comet assay 18 IX. Quantitative real time (qRT)-PCR 19 X. RNA-sequencing analysis 20 RESULTS 22 I. Molecular characterization of the rfx1 gene 22 II. Effects of rfx1 deletion on nuclei and septum formation 22 III. Virulence and toxin production 31 IV. Sexual development 31 V. Localization of GFP-tagged RFX1 proteins 35 VI. Sensitivity of rfx1 deletion strains to DNA-damage 35 VII. Accumulation of spontaneous DNA DSBs 38 VIII. Effects of rfx1 deletion on genome-wide transcription profiles 40 DISCUSSION 46 LITERATURE CITED 52 | - |
dc.format | application/pdf | - |
dc.format.extent | 3360961 bytes | - |
dc.format.medium | application/pdf | - |
dc.language.iso | en | - |
dc.publisher | 서울대학교 대학원 | - |
dc.subject | Fusarium graminearum | - |
dc.subject | Transcription factor | - |
dc.subject | RFX1 | - |
dc.subject | DNA double-strand break | - |
dc.subject | Cell division | - |
dc.subject | RNA sequencing | - |
dc.subject.ddc | 630 | - |
dc.title | Transcription factor RFX1 is critical for maintenance of chromosome integrity in Fusarium graminearum | - |
dc.title.alternative | Fusarium graminearum의 전사조절인자 RFX1의 기능연구 | - |
dc.type | Thesis | - |
dc.description.degree | Doctor | - |
dc.citation.pages | 62 | - |
dc.contributor.affiliation | 농업생명과학대학 농생명공학부 | - |
dc.date.awarded | 2014-08 | - |
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