2005 Fiscal Year Final Research Report Summary
Interactions of the retinoid X receptor with a RING protein and its effects on ubiquitylation and transcription
| Project/Area Number |
16570093
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Structural biochemistry
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| Research Institution | Gifu University |
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Principal Investigator |
OKANO Yukio Gifu University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (10177066)
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| Co-Investigator(Kenkyū-buntansha) |
MUTO Yoshinori Gifu University, School of Medicine, Professor, 医学部, 教授 (80190859)
YOSHIOKA Takashi Gifu University, Graduate School of Medicine, Research Associate, 大学院・医学系研究科, 助手 (20311699)
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| Project Period (FY) |
2004 – 2005
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| Keywords | RING-finger / retinoid X receptor (RXR) / Ubc13 / transactivation / ubiquitylation / Lys63 / dominant negative / FRET |
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| Research Abstract |
We previously reported the interaction of the RING-finger protein, RNF8, a binding partner of UBE2E2 (a ubiquitin-conjugating enzyme), with retinoid X receptor (RXR). Using the deletion mutants of RNF8 and RXR, the important domains in these proteins were examined by two-hybrid assay or in vitro pull-down assay. Wild-type RNF8 were distributed in nucleus, whereas an N-terminal deletion mutant (deltaN) and the RING-disrupted mutant (dominant negative) of RNF8 were observed in cytosole. Although RNF8 did not seem to act as an E3 for RXR ubiquitylation, it enhanced RXR transcriptional activity independently of a selective ligand, 9-cis retinoic acid. This transactivation effects were abolished with deltaN and dominant negative mutants of RNF8.
Furthermore, in collaboration with Dr.Timothy Thomson's group in Spain, following observations were made. Two-hybrid screening using UBC13 as a bait, we cloned the variant E2, UEV, and several RING-finger proteins including RNF8, KIAA00675, CHFR, KF1, and ZNRF2. UEV is known to form multi-ubiquitin chains via K63 but not ordinary K48. Autoubiquitylation of RNF8 was examined using wild-type ubiquitin (UbWT), and 3 types of K to R mutants of ubiquitin (UbK48,63R; UbK29,63R; UbK29,48R). In vitro ubiquitylation experiments in combination with the expressions of dominant negative UBC13 mutant (UBC13-C87A) and UbK29,48R, revealed ubiquitylation of RNF8 via K63 but not K48.
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Research Products
(14 results)
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[Journal Article] The RING finger protein, RNF8, interacts with retinoid X receptor α and enhances its transcription-stimulating activity.2004
Author(s)
Takano Y, Adachi S, Okuno M, Muto Y, Yoshioka T, Matsushima-Nishiwaki R, Tsurumi H, Ito K, Friedman.S.L, Moriwaki H, Kojima S, Okano Y
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Journal Title
J Biol Chem 279
Pages: 18926-18934
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Aurora-C kinase is a novel chromosomal passenger protein that can complement Aurora-B kinase function in mitotic cells.2004
Author(s)
Sasai K, Katayama H, Stenoien DL, Fujii S, Honda R, Kimura M, Okano Y, Tatsuka M, Suzuki F, Nigg EA, Earnshaw WC, Brinkley WR, Sen Subrata
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Journal Title
Cell Motil Cytoskelet 59
Pages: 249-263
Description
「研究成果報告書概要(欧文)」より
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