1999 Fiscal Year Final Research Report Summary
Identification of protein components in yeast mitochondrial nuclei.
| Project/Area Number |
09680697
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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 |
Cell biology
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| Research Institution | Yamaguchi University |
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Principal Investigator |
MIYAKAWA Isamu Yamaguchi University, Department of Biology, Associate Professor., 理学部, 助教授 (50136165)
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| Co-Investigator(Kenkyū-buntansha) |
宮川 勇 山口大学, 理学部, 助教授 (50136165)
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| Project Period (FY) |
1997 – 1999
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| Keywords | yeast / Saccharomyces cerevisiae / mitochondria / mitochondrial nuclei / mitochondrial nucleoids / DNA-binding protein |
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| Research Abstract |
1. We isolated mitochondrial (mt) nuclei from anaerobically-grown cells, respiratory deficient cells and respiration-inhibited cells. We found that these mt-nuclei contained significantly reduced amount of 67 kDa protein, compared with mt-nuclei from aerobically-grown cells.
2. The N-terminal amino acid sequence of the 67 kDa protein was determined. This protein was proved to be a carnitine acetyltransferase (CAT) that localized in mitochondria. Immunological assay using the antibody against CAT suggested association of CAT with mt-nuclei.
3. Behavior of mitochondria, mt-nuclei, microtubules and actin were investigated by fluorescence microscopy in the triangular yeast, Trigonopsis variabilis.
4. We demonstrated that Abf2p, a major DNA-binding protein of mitochondria, is tightly associated with mt-nuclei by Western blotting and immunofluorescence microscopy. We showed that SDS-DNA PAGE is an efficient for detection of Abf2p on gels.
5. We identified the YMN-1 antigen protein of mt-nuclei, which is recognized by YMN-1 monoclonal antibody, as dihydrolipoyl transsuccinylase (KE2). Enzyme assay suggested that α-ketoglutarate dehydrogenase complex (KGDC) cosedimented with mt-nuclei in sucrose-density gradient.
6. Protein components of mt-nuclei were separated by two-dimensional (2D) electrophoresis. We determined N-terminal amino acid sequence of 54 kDa, 52 kDa, 50 kDa, 38 kDa and 22 kDa proteins, respectively, which are involved in mt-nuclei.
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