2003 Fiscal Year Final Research Report Summary
Structural analyses of transcriptional regulation in stress response
| Project/Area Number |
13480222
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biophysics
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| Research Institution | Yokohama City University |
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Principal Investigator |
NISHIMURA Yoshifumi Yokohama City University, Graduate school of integrated Science, Professor, 総合理学研究科, 教授 (70107390)
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| Project Period (FY) |
2001 – 2003
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| Keywords | transcription factor / telomeres / DNA-binding protein / TRF1 / TRF2 / NMR / TFIIE / Zn binding domain |
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| Research Abstract |
The zinc finger domain in the large subunit of TFIIE is phylogenetically conserved and is essential for transcription. We have determined its NMR structure. It consists of one alpha helix and six beta strands showing novel features distinct from previously determined zinc-binding structures. We created point mutants in this domain and examined their binding abilities to other general transcription factors as well as their transcription activities. Interesting functional asymmetry of Zn^<2+>-ligand mutants was observed: the N-terminal two mutants remained approximately 20% activity on a supercoiled template. CD and NMR studies showed that those two equilibrate mainly with the random coil structure, although all four mutants possessed partially folded characteristic structures coordinating Zn^<2+>-. And also, highly conserved D 164 mutants (D 164A and D 164K) were found to increase transcription.
In addition, we have determined the solution structure of the DNA binding domain of hTRF2 bound to a telomeric double-stranded DNA with the sequence of GTTAGGGTTAGGG and compared it with the corresponding DNA complex structure of hTRF1. Telomeres are the ends of eukaryotic linear chromosomes consisting of repetitive G-rich sequence and telomeric repeat binding factors. In mammalian telomeres, TRF1 and TRF2 bind to double-stranded telomeric DNA. Both contain a central TRF-homology (TRFH) domain and a C-terminal DNA binding domain. Both DNA-bound structures are very close to each other, however, small but significant structural differences are observed. Based on the present structure of hTRF2 we could make several mutants of hTRF2, which have a stronger binding ability to telomeric DNA rather than the wild type.
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Research Products
(21 results)
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[Journal Article] Selective dissociation of non-covalent bonds in biological molecules by laser spray.2004
Author(s)
Takamizawa, A., Ito, Y., Osawa, R., Iwasaki, N., Nishimura, Y., Akashi, S., Hiraoka,K
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Journal Title
Journal of mass Spectrometetry Vol,39
Pages: 1053-1058
Description
「研究成果報告書概要(和文)」より
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[Journal Article] A Novel zinc finger structure in the large Subunit of human general transcription factor TFIIE.2004
Author(s)
Okuda, M., Tanaka, A., Arai, Y., Satoh, M., Okamura, H., Nagadoi, A., Hanaoka, F., Ohkuma, Y., Nishimura, Y.
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Journal Title
The Journal of Biological Chemistry 279
Pages: 51395-51403
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Selective dissociation of non-covalent bonds in biological molecules by laser spray.2004
Author(s)
Takamizawa, A., Ito, Y., Osawa, R, Iwasaki, N., Nishimura, Y., Akashi, S., Hiraoka, K.
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Journal Title
Journal of mass Spectrometetry 39
Pages: 1053-1058
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Crystallization and preliminary X-ray diffraction studies on the DNA-binding domain of the transcriptional activator protein PhoB from escherichia coli.2002
Author(s)
Shindoh, K., Maenaka, K., Akiba, T., Okamura, H., Nishimura, Y., Makino, K., Shirakihara,Y
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Journal Title
Acta Crystallography Vol,58
Pages: 1862-1864
Description
「研究成果報告書概要(和文)」より
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[Journal Article] A Novel zinc finger structure in the large subunit of human general transcription factor TFIIE.2002
Author(s)
Okuda, M., Tanaka, A., Arai, Y., SAto, M., Okamura, H., Nagadoi, A., Hanaaoka, F., Ohkuma, Y., Nishimura,Y
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Journal Title
The Journal of Biological Chemistry Vol,279
Pages: 51395-51403
Description
「研究成果報告書概要(和文)」より
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[Journal Article] Crystallization and preliminary X-ray diffraction studies on the DNA-binding domain of the transcriptional activator protein PhoB from Escherichia coli.2002
Author(s)
Shindoh, K., Maenaka, K., Akiba, T., Okamura, H., Nishimura, Y., Makino, K., Shirakihara, Y.
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Journal Title
Acta Crystallography 58
Pages: 1862-1864
Description
「研究成果報告書概要(欧文)」より
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