| Project/Area Number |
15014213
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas
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| Allocation Type | Single-year Grants |
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| Review Section |
Biological Sciences
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| Research Institution | THE UNIVERSITY OF TOKYO |
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Principal Investigator |
ITO Takashi THE UNIVERSITY OF TOKYO, GRADUATE SCHOOL OF FRONTIER SCIENCES, PROFESSOR, 大学院新領域創成科学研究科, 教授 (90201326)
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| Co-Investigator(Kenkyū-buntansha) |
SUMMOTO Hidekl KYUSHUUNIVERSITY, MEDICAL INSTITUTE OF BIO-REGULATION, PROFESSOR, 生体防御医学研究所, 教授 (30179303)
OTA Kazuhisa THE UNIVERSITY OF TOKYO, GRADUATE SCHOOL OF FRONTIER SCIENCES, RESEARCH ASSOCIATE, 大学院新領域創成科学研究科, 助手 (00322727)
YAMADA Yoichi KANAZAWA UNIVERSITY, GRADUATE SCHOOL OF NATURAL SCIENCES, RESEARCH ASSOCIATE, 大学院自然科学研究科, 助手 (30377402)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥50,600,000 (Direct Cost: ¥50,600,000)
Fiscal Year 2004: ¥25,600,000 (Direct Cost: ¥25,600,000)
Fiscal Year 2003: ¥25,000,000 (Direct Cost: ¥25,000,000)
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| Keywords | PROTEIN-PROTEIN INTERACTION / YEAST 2-HYBRID SYSTEM / PB1 DOMAIN / TAP TAG / UBIQUITIN / MASS SPECTROMETRY / DNA METHYLATION / FULL LENGTH cDNA / 蛋白質間相互作用 / 2ハイブリッド法 / 質量分析法 / ユビキチン化 / 蛋白質DNA相互作用 / 1ハイブリッド法 / 相互作用ドメイン / 蛋白質核酸相互作用 / 転写開始点 / 出芽酵母 |
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| Research Abstract |
1)We performed a comprehensive two-hybrid analysis for the budding yeast proteome and made the data available.
2)We utilized the two-hybrid clone resource to conduct genome-wide screens for chromation bariier proteins as well as transcriptional activators.
3)We developed two-hybrid footprinting, dual-bait reverse two-hybrid method, and accelerated phylogenetic footprinting to conduct interaction-targeting or targeted disruption of proteinprotein interactions.
4)We conducted a search for rules for protein-protein interactions by means of data-mining using the association rule discovery.
5)We compared yeast and mammalian systems of mutually interacting proteins including PB1-domain proteins and revealed two-different modes of target recognition by PB1 domains, enabling us to predict the PB1 interactions.
6)We developed a novel TAP (Tandern Affinity Purification) tag that am be used with crosslinking agents for efficient interaction profiling.
7)We developed a method of PAP (Parallel Affinity Purification) to enrich ubiquitinated proteins for degradome profiling.
8)We developed a methylation-dependent one-hybrid system to examine interactions between methylated DNA and proteins.
9)We developed HM-PCR for allelic methylation analysis and applied it to a comprehensive analysis of CpG islands on human chromosome 21.
10)We developed a method for chimaerization-mediated activation of transcription factors and applied it to decipher a gene network regulating multidrug resistance of the budding yeast
11)We systematically analyzed budding yeast full-length cDNA clones to reveal transcription starting sites for-4,000 genes.
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