2012 Fiscal Year Final Research Report
Establishment of simultaneous and quantitative assay for DNAbinding and transcriptional activity of glucocorticoid receptor.
| Project/Area Number |
23770169
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Biophysics
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| Research Institution | Hokkaido University |
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Principal Investigator |
MIKUNI Shintaro 北海道大学, 大学院・医学研究科, 特任助教 (40435954)
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| Project Period (FY) |
2011 – 2012
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| Keywords | タンパク質 / 核酸の構造・動態・機能 / 転写因子 / バイオイメージング / 分子間相互作用 |
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| Research Abstract |
Regulation of transcriptional activity could be initiated by associating the transcriptional factors with its specific sequence of genomic DNA. However, the direct relationship between “the affinity for DNA” and “transcriptional activity” of transcriptional factors is not clarified. To clarify this relationship, we tried to establish the simultaneous and quantitative assay for DNA binding and transcriptional activity of glucocorticoid receptor (GR), which is one of transcriptional factors, using fluorescence cross-correlation spectroscopy (FCCS). As a result, the dissociation constant between the purified GR fused with EGFP and the Alexa647-labeled GR specific sequences (glucocorticoid response element, GRE) could be determined as 0.63 μM, and sequence-specificity of GR could be confirmed by FCCS. Moreover, the dissociation constant between the purified RNA-specific binding protein fused withmCherry (red fluorescent protein) and Alexa488-labeled RNA was also determined as 143 nM by FCCS.
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Research Products
(6 results)
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[Journal Article] Virus-like particles with removable1 cyclodextrins enable glutathione-triggered drug release in cells.2013
Author(s)
Niikura K, Sugimura N, Musashi Y, Mikuni S, Matsuo Y, Kobayashi S, Nagakawa K, Takahara S, Takeuchi C, Sawa H, Kinjo M, Ijiro K.
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Journal Title
Mol Biosyst.
Volume: 9(3)
Pages: 501-7
DOI
Peer Reviewed
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[Journal Article] Rab6a releases LIS1 from a dynein idling complex and activates dynein for retrograde movement.
Author(s)
Yamada M, Kumamoto K, Mikuni S, Arai Y, Kinjo M, Nagai T, Tsukasaki Y, Watanabe TM, Fukui M, Jin M, Toba S, Hirotsune S.
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Journal Title
Peer Reviewed
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