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Roles of the Histone Acetyltransferase Rtt109 in Genome Integrity Maintenance of Magnaporthe oryzae : 벼 도열병균 유전체 보전에 관여하는 히스톤 라이신 아세틸화 효소 Rtt109의 역할

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dc.contributor.advisor이용환-
dc.contributor.author권서문-
dc.date.accessioned2017-07-14T06:44:05Z-
dc.date.available2017-07-14T06:44:05Z-
dc.date.issued2015-02-
dc.identifier.other000000026476-
dc.identifier.urihttps://hdl.handle.net/10371/125898-
dc.description학위논문 (석사)-- 서울대학교 대학원 : 농생명공학부, 2015. 2. 이용환.-
dc.description.abstractPreservation of genome integrity is undoubtedly a fundamental function of life, ensuring the survival and propagation of specific genetic assemblages selected over time. The fungal-specific histone acetyltransferase Rtt109 is implicated in various chromatin functions influencing genome integrity and plasticity. In particular, Rtt109 plays a crucial role in nucleosome assembly associated with DNA replication and repair processes. This thesis explores the implications of Rtt109-mediated genome integrity maintenance in the plant pathogen Magnaporthe oryzae, regarding pathogenicity and mycological development. Deletion of Rtt109 led to a marked decrease in mycelial growth rate and a concomitant growth reduction in planta. The growth defect seems to be linked to the elevated level of DNA damage constitutively suffered by the mutant, even in the absence of genotoxins. Accordingly, expression of DNA damage repair genes and stress-related genes were increased. The deletion mutant was also rendered hypersensitive to various forms of genotoxic stress including ROS, which is integral to fungal morphogenesis and the plant defense response. However, appressorium morphogenesis was unaffected by the absence of Rtt109 and the extent of growth reduction in planta was not beyond the innate mycelial growth defect. Therefore it is considered that the level of ROS and genotoxic stress experienced by M. oryzae during interaction with the host plant is not sufficient to hinder infectious development, even in the vulnerable state without Rtt109. Furthermore, it can be inferred that the intrinsic protective mechanisms together with the up-regulation of DNA damage and stress response genes may be sufficient to protect the mutant throughout the lifecycle. In conclusion, Rtt109-mediated genome preservation is required for normal growth and genotoxic stress resistance in M. oryzae. Although Rtt109 is not directly involved in pathogenicity, it is expected to have important contributions to genome regulation and evolution.-
dc.description.tableofcontentsABSTRACT i
CONTENTS iii

INTRODUCTION 1

MATERIALS AND METHODS 9
I. Strains and growth conditions 9
II. Targeted deletion of Rtt109 and genetic complementation 9
III. Western blot analysis of H3K56 acetylation 10
IV. Phenotypic complementation of Saccharomyces cerevisiae rtt109Δ mutant 10
V. Cell wall staining and hyphal cell length measurements - 11
VI. Genotoxin and ROS sensitivity tests 11
VII. Neutral comet assay 12
VIII. Pathogenicity tests 12
IX. Observations on asexual and pre-penetration
development 13
X. qRT-PCR and RNA sequencing 14

RESULTS 18
I. Identification of Rtt109 ortholog in Magnaporthe oryzae- 18
II. Evidence of M. oryzae Rtt109 functionality as a H3K56 acetyltransferase 21
III. Deletion of Rtt109 leads to reduced mycelial growth and increases DNA double strand breaks 24
IV. Δrtt109 is hypersensitive to genotoxins and reactive oxygen species 28
V. Pathogenicity is compromised in Δrtt109 30
VI. Deletion of Rtt109 leads to gross changes in expression of DNA metabolism and repair genes 35

DISCUSSION 40
I. M. oryaze requires Rtt109 for genome stability and genotoxin resistance 40
II. Rtt109 is required for full tolerance to ROS but is dispensable for pre-penetration development and pathogenicity 42
III. DNA damage and repair in the rice blast fungus 46
IV. Clues from the expression analysis 47
V. Conclusion and suggestions for further research 48

BIBLIOGRAPHY 52
ABSTRACT IN KOREAN 60
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dc.formatapplication/pdf-
dc.format.extent1799889 bytes-
dc.format.mediumapplication/pdf-
dc.language.isoen-
dc.publisher서울대학교 대학원-
dc.subjectHistone acetyltransferase-
dc.subjectMagnaporthe oryzae-
dc.subject.ddc630-
dc.titleRoles of the Histone Acetyltransferase Rtt109 in Genome Integrity Maintenance of Magnaporthe oryzae-
dc.title.alternative벼 도열병균 유전체 보전에 관여하는 히스톤 라이신 아세틸화 효소 Rtt109의 역할-
dc.typeThesis-
dc.contributor.AlternativeAuthorSeomun Kwon-
dc.description.degreeMaster-
dc.citation.pagesiv, 64-
dc.contributor.affiliation농업생명과학대학 농생명공학부-
dc.date.awarded2015-02-
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