| Project/Area Number |
18591018
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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 |
Endocrinology
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| Research Institution | Nagoya University |
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Principal Investigator |
KANOU Yasuhiko Nagoya University, Research Instiiut of Environmental Medicine, Assistant Professor (50252292)
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| Co-Investigator(Kenkyū-buntansha) |
HAYASHI Yoshitaka Nagoya University, Research Inslilute of Fnvimnmental Medicine, Associate professor (80420363)
TAKAGISHI Yoshiko Nagoya University, Research Institute of Envimnmenlal Medicine, Assistant professor (50024659)
MURATA Yoshiharu Nagoya University, Research Institute of Environmental Medicine, Professor (80174308)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,670,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥270,000)
Fiscal Year 2007: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2006: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | Resistance to thyroid hormone / thyroid hormone receptors / co-activator / Yeast two-hybrid system / cDNA / cloning |
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| Research Abstract |
There is a case where any mutations are not found in two types of thyroid hormone receptor(TR) in the resistance to thyroid hormone. We hypothesized that a specific coactivator for thyroid hormone receptor b existed. This coactivator may have any abnormality in the resistance to thyroid hormone. We tried to identify the cDNA for the specific protein. Yeast two-hybrid system was applied to screen a human liver cDNA library. We found two cDNA clones, No.60 and 93, as a candidate of TR-specific coactivator.
1. A clone 60 was a fragment derived from 3 kinds of mRNA. The full-length cDNAs for these mRNAs were cloned. The deduced amino acid sequences showed that proteins had nuclear transport signals.
2. In amino acid sequences of clone 60 proteins, there was a high homology sequence for another nuclear protein.
3. To determine whether a clone 60 protein binds to TRb, pull-down assay was occurred. TRb and a clone 60 protein were expressed as a GST-fusion protein and His-tag fusion protein, respectively. Both fusion protein were mixed and precipitated with specific antibodies. Western blotting showed both fusion proteins bound in vitro.
4. A clone 93 could not be expressed as His-tag fusion protein in vitro. We are advancing the analysis of the clone 60 proten further continuously.
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