2007 Fiscal Year Final Research Report Summary
Molecular mechanism of prion propagation revealed by yeast prion as a model
| Project/Area Number |
17370034
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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 |
Structural biochemistry
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| Research Institution | The University of Tokyo |
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Principal Investigator |
TAGUCHI Hideki The University of Tokyo, Graduate School of Frontier Sciences, Associate Professor (40272710)
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| Project Period (FY) |
2005 – 2007
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| Keywords | prion / protein folding / chaperones / Saccharomyces cerevisiae / amyloid / protein / protein aggregation |
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| Research Abstract |
Prions are infectious proteins; an abnormal form of the prion protein causes an auto-catalytic conversion of a normal (soluble) form of prion protein, which has the same primary structure as the abnormal one, to the abnormal insoluble form, and results in accumulation of infectious insoluble prion particles. In many cases, the particles are observed as amyloid fiber-like structures, which are called ‘prion-fibrils'. This concept originated from the studies of mammalian neurodegenerative diseases, but spread to include other proteinous genetic elements from yeast Saccharomyces cerevisiae. In particular, the prion-inducing fragment of yeast Sup35, a determinant of the yeast prion-like [PSI^+], is comprised of a glutamine/asparagine-rich N-terminal and medium domains of Sup35, and is a valuable model protein for studying the mechanism of prion-fiber formation.
Using the yeast prion as a model, we have previously developed a system to observe the growth of individual prion fibrils directly.
… More
As a first objective in this study, we have further expanded the imaging using total internal reflection fluorescent microscopy (TIRFM). By using the TIRFM system, we succeeded in monitoring live imaging of individual Sup35 fibrils elongation. Statistical analysis of rate of the fibril growth provides the kinetic information, and then a novel insight into the mechanism of fibril growth.
Second, we directly monitored the dynamics of the aggregates in living cells using an on-chip single-cell cultivation system as well as fluorescence correlation spectroscopy (FCS). Single-cell imaging revealed that the visible foci of yeast prion Sup35 fused with GFP are dispersed throughout the cytoplasm during cell growth, but retain the prion phenotype. FCS showed that [PSI^+] cells, irrespective of the presence of foci, contain diffuse oligomers, which are transmitted to their daughter cells. Single-cell observations of the oligomer-based transmission provide a link between previous in vivo and in vitro analyses of the prion and shed light on the relationship between the protein conformation and the phenotype. Less
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[Journal Article] Bimodal protein solubility distribution revealed by an aggregation analysis of the entire ensemble of Escherichia coli proteins2009
Author(s)
Niwa, T., Ying, B. -W., Saito, K, Jin, W. Z., Takada, S., Ueda, T. Taguchi, H.
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Journal Title
Proc. Natl. Acad. Sci. U.S.A. (印刷中)(掲載確定)
Description
「研究成果報告書概要(和文)」より
Peer Reviewed
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[Journal Article] Bimodal protein solubility distribution revealed by an aggregation analysis of the entire ensemble of Escherichia coli proteins.2009
Author(s)
Niwa, T., Ying, B. -W., Saito, K., Jin, W. Z., Takada, S., Ueda, T, Taguchi, H.
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Journal Title
Proc. Natl. Acad. Sci. U.S.A. (in press)
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Kinetic analysis of conformational changes of GroEL based on the fluorescence of tyrosine 506.2008
Author(s)
Hosono, K, Ueno, T., Taguchi, H., Motojima, F, Zako, T., Yoshida, M., Funatsu, T.
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Journal Title
Protein J. 27
Pages: 461-468
Description
「研究成果報告書概要(和文)」より
Peer Reviewed
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[Journal Article] Single molecule imaging of full protein synthesis by immobilized ribosomes.2008
Author(s)
Uemura, S., Iizuka, R., Ueno, T., Shimizu, Y, Taguchi, H., Ueda, T., Puglisi, J., Funatsu, T.
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Journal Title
Nucleic Acids Research 36
Pages: 70-74
Description
「研究成果報告書概要(和文)」より
Peer Reviewed
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[Journal Article] Dynamics of yeast prion aggregates in single living cells.2006
Author(s)
Kawai-Noma, S., Ayano, S., Pack, C-G., Kinjo, M., Yoshida, M., Yasuda, K., Taguchi, H.
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Journal Title
Genes to Cells 11
Pages: 1085-1096
Description
「研究成果報告書概要(和文)」より
Peer Reviewed
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