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Post-doctoral Fellow Center
for Structural Biology
Phone: (203) 432-5625
Email: fujii@csb.yale.edu |
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Post-doctoral Fellow Center
for Structural Biology
Phone: (203) 432-5625
Email: fujii@csb.yale.edu |
Structural studies of Mitochondrial Ribosome
Summary:
In contrast to the bacterial and eukaryotic translation systems, mitochondrial translation system has unique features: (a) use of non-universal codons, (b) use of abnormal tRNAs different from normal clover structure, (c) use of only 22 tRNAs to read 61 codons, and (d) a much larger mass ratio of protein to rRNA.
From these particular features, three main questions arise: (1) How translation using non-universal codons with abnormal tRNAs occurs? (2) How are 61 codons read using only 22 tRNAs? (3) What are the functions of ribosomal proteins that replace rRNAs? To elucidate these questions, we are experimenting to obtain crystals of mitochondorial ribosome for structure determination.
The purification of mitoribosome has been challenging due to their low abundance (2~3% of total mitochondrial proteins). To get high quality crystals of ribosome, the most important thing is the homogeneity of the sample. To overcome these difficulties (quantity and quality), we are planning two experimental designs using yeast as a target species.
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Publication:
Commentary on this work:
PAPERS RELEVANT TO YOUR WORK Friedberg EC, Fischhaber PL, Kisker C (2001) Error-Prone DNA Polymerases:
Novel Structures and the Benefits of Infidelity. Cell
107: 9-12. (Minireview)
Beard WA, Wilson SH (2001) DNA Lesion Bypass Polymerases Open Up. Structure
9: 759-764. (Minireview)
Ellenberger T, Silvian LF (2001) The anatomy of infidelity. Nat
Struct Biol 8: 827-828. (News & Views)
(2) Fujii, Y., Shimizu, T., Kyogoku, Y., Toda, T., Yanagida, M. & Hakoshima, T., (2000). Structural basis for the DNA recognition diversity of bZIP transcription factors revealed by the crystal structure of Pap1/DNA complex. Nature Structural Biology, 7, 889-893.
(3) Fujii, Y., Kusumoto, M., Shimizu, T., Kyogoku, Y., Taniguchi, T. & Hakoshima, T., (1999). Crystal structure of IRF /DNA complex reveals novel DNA recognition and cooperative binding to a tandem repeat of core sequences. The EMBO Journal, 18, 5028-5041.
(4) Shimizu, T., Fujii, Y.& Hakoshima, T., (1999). A structural view of intracellular signaling pathways of cytokines. Cell Technology, 18, 1474-1480.
(5) Fujii, Y., Ohira, T., Kyogoku, Y., Toda, T., Yanagida, M. & Hakoshima, T., (1998). Crystallographic Characterization of Pap1-DNA complex. Acta Crystallographica Section D, 54, 1014-1016.
(6) Kusumoto, M., Fujii, Y., Tsukuda, Y., Ohira, T., Kyogoku, Y., Taniguchi, T. & Hakoshima, T., (1998). Crystallographic Characterization of DNA-binding Domain of Interferon Regulatory Factor-2 Complexed with DNA. Journal of Structural Biology, 121, 363-366.
(7) Kuge, M., Fujii, Y., Shimizu, T., Hirose, F., Matsukage, A. & Hakoshima, T., (1997). Use of fusion protein to obtain crystals suitable for X-ray analysis: Crystallization of a GST-fused protein containing the DNA-binding domain of DNA replication-related element-binding factor, DREF. Protein Science, 6, 1783-1786.