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Global analysis of Yor289w interactome in Saccharomyces cerevisiae : 효모에서 Yor289w interactome의 분석

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dc.contributor.advisor허원기-
dc.contributor.author김지원-
dc.date.accessioned2017-07-19T09:07:28Z-
dc.date.available2017-07-19T09:07:28Z-
dc.date.issued2015-02-
dc.identifier.other000000025108-
dc.identifier.urihttps://hdl.handle.net/10371/131581-
dc.description학위논문 (석사)-- 서울대학교 대학원 : 생명과학부, 2015. 2. 허원기.-
dc.description.abstractIdentifying protein-protein interactions is essential for understanding protein function, since most cellular processes are mediated by interaction of proteins. Therefore, by identifying the interactome of a protein, we could determine the function of the protein. In this study, we conducted a genome-wide bimolecular fluorescent complementation (BiFC) assay in Saccharomyces cerevisiae to detect the protein-protein interactions of Yor289w and thus identify its functional role. A pRPL7B-VN-YOR289W strain and strains from a VC fusion library were mated and BiFC signals were visualized. Subsequent to BiFC screening and elimination of false positive signals, a total 56 candidates exhibited interactions with Yor289w. In the analysis of interactome, we found candidates clustered into processes related to translation initiation and nucleobase-containing small molecule metabolic processes. In addition we identified Yor289w protein expression increased in response to unfolded protein response (UPR), and that this increase was dependent on Hac1 and Ire1, which are key factors in UPR. This indicates Yor289w expression is under control of Hac1 and Ire1. On both genome-wide screening and expression data, we concluded that Yor289w might be involved in translational regulation under UPR conditions.-
dc.description.tableofcontentsABSTRACT…………………………………………………………………………………………… … …i
CONTENTS………………………………………………………………………………………………....ii
LIST OF FIGURES iv
LIST OF TABLES v
LIST OF ABBREVIATIONS vi

I. INTRODUCTION 1

II. MATERIALS AND METHODS 4
1. Yeast strains and growth media 4
2. Transformation of yeast cells for YOR289W analysis 4
3. Microscopic analysis 8
4. Western blot analysis 8
5. Real-Time PCR analysis 9

III. RESULTS 10
1. Identification of Yor289w expression and cellular localization 10
2. Identification of the Yor289w interactome 12
2-1. Genome-wide mating and selection for microscopic analysis 12
2-2. Genome-wide BiFC screening of Yor289w and elimination of self-assembly candidates 12
2-3. Functional analysis of the Yor289w interactome 17
3. Yor289w protein expression under stress conditions 26
3-1. Dithiothreitol and tunicamycin treatment induces expression of Yor289w 26
3-2. The Hac1 transcription factor controls the expression of YOR289W under UPR inducing conditions 27

IV. DISCUSSION 31

V. REFERENCE 34

국문초록 39
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dc.formatapplication/pdf-
dc.format.extent3398682 bytes-
dc.format.mediumapplication/pdf-
dc.language.isoen-
dc.publisher서울대학교 대학원-
dc.subjectYOR289W-
dc.subjectProtein-protein interaction-
dc.subjectBiFC assay-
dc.subjectSaccharomyces cerevisiae-
dc.subjectUnfolded protein response (UPR)-
dc.subject.ddc570-
dc.titleGlobal analysis of Yor289w interactome in Saccharomyces cerevisiae-
dc.title.alternative효모에서 Yor289w interactome의 분석-
dc.typeThesis-
dc.contributor.AlternativeAuthorKim Jiwon-
dc.description.degreeMaster-
dc.citation.pages40-
dc.contributor.affiliation자연과학대학 생명과학부-
dc.date.awarded2015-02-
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