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Study of DNA-Binding Proteins of Rat Liver Mitochondria

  • Biochemistry
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Abstract

Acid-soluble proteins able to form DNA-protein complexes in the presence of physiological concentration of NaCl were isolated from rat liver mitochondria. Electrophoretic analysis of these proteins in 15% polyacrylamide gel showed that mitochondrial acid-soluble proteins include of approximately 20 polypeptides with molecular weight of 10–120 kDa. The fraction of acid-soluble proteins can be separated into basic and acidic proteins by chromatography on DEAE cellulose. Some of acidic proteins are tightly bound to the basic proteins and can be separated from them in the presence of 5 mM dithiothreitol. It is discovered that the fraction of acidic proteins contains proteases (including DNA-activated ones), which cleave different polypeptides of the basic proteins with different efficiency. Possibly, mitochondrial DNA-binding proteins and DNA-activated proteases are involved in the regulation of structural organization and functional activity of mitochondrial DNA.

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REFERENCES

  • Alam, T.I., Kanki, T., Muta, T., Ukaji, K., Abe, Y., Nakayama, H., Takio, K., Hamasaki, N., and Kand, D., Human Mitochondrial DNA Is Packaged with TFAM, Nucleic Acid Res., 2003, vol. 31, pp. 1640–1645.

    Article  PubMed  Google Scholar 

  • Anderson, S., Bankier, A.T., Barrel, B.G., de Bruijn, M.H.L., Coulson, A.R., Drouin, A.J., Eperon, I.C., Nierlich, D.P., Roe, B.A., Sanger, F., Schreier, P.H., Smith, A.J.H., Staden, R., and Young, I.G., Sequence and Organisation of the Human Mitochondrial Genome, Nature, 1981, vol. 290, pp. 457–465.

    Article  PubMed  Google Scholar 

  • Bibb, M.J., Van Etten, R.A., Wright, C.T., Walberg, M.W., and Clayton, D.A., Sequence and Gene Organization of Mouse Mitochondrial DNA, Cell, 1981, vol. 26, pp. 167–180.

    Article  PubMed  Google Scholar 

  • Blobel, G. and Potter, V.R., Nuclei from Rat Liver: Isolation Method That Combines Purity with High Yield, Science, 1966, vol. 154, pp. 1662–1665.

    PubMed  Google Scholar 

  • Bonner, J., Chalkley, G.R., Dahmus, M., Fambrough, D., Fujimura, F., Huang, R.-C.C., Huberman, J., Jensen, R., Marushige, K., Ohlenbush, H., Olivera, B., and Widholm, J., Isolation and Characterization of Chromosomal Nucleoprotein, in Methods Enzymol., Grossman, L. and Moldave, K., Eds., New York: Academic, 1968, vol. 12B, pp. 3–64.

    Google Scholar 

  • Bota, D.A. and Davies, K.J.A., Protein Degradation in Mitochondria: Implications for Oxidative Stress, Aging and Disease: A Novel Etiological Classification of Mitochondrial. Proteolytic Disorders, Mitochondrion, 2001, vol. 1, pp. 33–49.

    Article  PubMed  Google Scholar 

  • Caron, F., Jacq, C., and Rouviere-Yaniv, J., Characterization of a Histone-Like Protein Extracted from Yeast Mitochondria, Proc. Natl. Acad. Sci. U.S.A., 1979, vol. 76, pp. 4265–4269.

    PubMed  Google Scholar 

  • Cortopassi, G.A., Shibata, D., Soong, N.-W., and Arnheim, N., A Pattern of Accumulation of a Somatic Deletion of Mitochondrial DNA in Aging Human Tissues, Proc. Natl. Acad. Sci. U.S.A., 1992, vol. 89, pp. 7370–7374.

    PubMed  Google Scholar 

  • Fisher, R.P., Lisowsky, T., Breen, G.A.M., and Clayton, D.A., A Rapid, Efficient Method for Purifying DNA-Binding Proteins, J. Biol. Chem., 1991, vol. 266, pp. 9153–9160.

    PubMed  Google Scholar 

  • Fu, G.K. and Markovitz, D.M., The Human Lon Protease Binds to Mitochondrial Promoters in a Single-Stranded, Site-Specific, Strand-Specific Manner, Biochemistry, 1998, vol. 37, pp. 1905–1909.

    Article  PubMed  Google Scholar 

  • Garrido, N., Griparic, L., Jokitalo, E., Wartiovaara, J., Van der Bliek, A.M., and Spelbrink, J.N., Composition and Dynamics of Human Mitochondrial Nucleoids, Mol. Biol. Cell, 2003, vol. 14, pp. 1583–1596.

    Article  PubMed  Google Scholar 

  • Hnilica, L.S., Methods for Analysis of Histones, in Methods Enzymol., O'Malley, B.,W. and Hardman, J.G., Eds., New York: Academic, 1975, vol. 40E, pp. 102–138.

    Google Scholar 

  • Kazakova, T.B., Mel'nikova, M.P., and Sverdlov, E.D., Isolation and Characterization of DNA Fragments Bound to Mitochondrial Membrane Proteins, Mol. Biol., 1977, vol. 11, no.5, pp. 1029–1037.

    Google Scholar 

  • Kelleher, P.J. and Juliano, P.L., Detection of Proteases in Polyacrylamide Gels Containing Covalently Bound Substrates, Anal. Biochem., 1984, vol. 136, pp. 470–475.

    Article  PubMed  Google Scholar 

  • Koc, E.C. and Spremulli, L.L., RNA-Binding Proteins of Mammalian Mitochondria, Mitochondrion, 2003, vol. 2, pp. 277–291.

    Article  PubMed  Google Scholar 

  • Kutsyi, M.P. and Gaziev, A.I., Gamma Irradiation Or Hydrocortisone Treatment of Rats Increases the Proteinase Activity Associated with Histones of Thymus Nuclei, Radiat. Res., 1994, vol. 140, pp. 221–229.

    PubMed  Google Scholar 

  • Kutsyi, M.P., Kuznetsova, E.A., Gulyaeva, N.A., and Gaziev, A.I., Effect of γ-Radiation and Mitochondrial Apoptogenic Factors on Activity of Nuclear Proteases, Radiats. Biol. Radioekol., 2002, vol. 42, no.4, pp. 357–363.

    Google Scholar 

  • Kutsyi, M.P., Zakrzhevskaya, D.T., and Gaziev, A.I., Hydrocortisone and Partial Hepatectomy Activate the Proteinase Associated with Histones, Biokhimiya, 1992, vol. 57, no.10, pp. 1548–1553.

    Google Scholar 

  • Laemmli, U.K., Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4, Nature, 1970, vol. 227, pp. 680–685.

    Article  PubMed  Google Scholar 

  • Loewenstein, J., Scholte, H.R., and Wit-Peeters, E.M., A Rapid and Simple Procedure To Deplete Rat-Liver Mitochondria of Lysosomal Activity, Biochim. Biophys. Acta, 1970, vol. 223, pp. 432–436.

    PubMed  Google Scholar 

  • Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J., Protein Measurement with the Folin Phenol Reagent, J. Biol. Chem., 1951, vol. 193, pp. 265–275.

    Google Scholar 

  • Marcelino, L.A. and Thilly, W.G., Mitochondrial Mutagenesis in Human Cells and Tissues, Mutat. Res., 1999, vol. 434, pp. 177–203.

    PubMed  Google Scholar 

  • Mignotte, B., Barat, M., and Mounolou, J.-C., Characterization of a Mitochondrial Protein Binding to Single-Stranded DNA, Nucleic Acids Res., 1985, vol. 13, pp. 1703–1716.

    PubMed  Google Scholar 

  • Miyakawa, I., Sando, N., Kawano, S., Nakamura, S., and Roiwa, N., Isolation of Morphologically Intact Mitochondrial Nucleoids from the Yeast Saccharomyces cerevisiae, J. Cell Sci., 1987, vol. 88, pp. 431–439.

    PubMed  Google Scholar 

  • Olszewska, E. and Tait, A., Mitochondrial Chromatin in Paramecium Aurelia, Mol. Gen. Genet., 1980, vol. 178, pp. 453–457.

    Article  PubMed  Google Scholar 

  • Ozawa, T., Genetic and Functional Changes in Mitochondria Associated with Aging, Physiol. Rev., 1997, vol. 77, pp. 425–464.

    PubMed  Google Scholar 

  • Pavco, P.A. and Van Tuyle, G.C., Purification and General Properties of the DNA-Binding Protein (P16) from Rat Liver Mitochondria, J. Cell Biol., 1985, vol. 100, pp. 258–264.

    Article  PubMed  Google Scholar 

  • Pedersen, P.L., Greenawalt, J.W., Reynafarje, B., Hullihen, J., Decker, G.L., Soper, J.W., and Bustamente, E., Preparation and Characterisation of Mitochondria and Submitochondrial Particles of Rat Liver and Liver-Derived Tissues, in Methods in Cell Biology, Prescott, D.M., Ed., New York: Academic, 1978, vol. 20, pp. 411–481.

    Google Scholar 

  • Potter, D.A., Fostel, J.M., Berninger, M., Pardue, M.L., and Cech, T.R., DNA-Protein Interactions in the Drosophila melanogaster Mitochondrial Genome as Deduced from Trimethylpsoralen Crosslinking Patterns, Proc. Natl. Acad. Sci. U.S.A., 1980, vol. 77, pp. 4118–4122.

    PubMed  Google Scholar 

  • Richter, C., Park, J.-W., and Ames, B.N., Normal Oxidative Damage to Mitochondrial and Nuclear DNA Is Extensive, Proc. Natl. Acad. Sci. U.S.A., 1988, vol. 85, pp. 6465–6467.

    PubMed  Google Scholar 

  • Richter, C., Reactive Oxygen and DNA Damage in Mitochondria, Mutat. Res., 1992, vol. 275, pp. 249–255.

    PubMed  Google Scholar 

  • Suter, M. and Richter, C., Fragmented Mitochondrial DNA Is the Predominant Carrier of Oxidized DNA Bases, Biochemistry, 1999, vol. 38, pp. 459–464.

    Article  PubMed  Google Scholar 

  • Takamatsu, C., Umeda, S., Ohsato, T., Ohno, T., Abe, Y., Fukuoh, A., Shinagawa, H., Hamasaki, N., and Kang, D., Regulation of Mitochondrial D-Loops by Transcription Factor A and Single-Stranded DNA-Binding Protein, EMBO Rep., 2002, vol. 18, pp. 451–456.

    Article  Google Scholar 

  • Van Tuyle, G.C. and Kalf, G.F., Isolation of Membrane-DNA-RNA Comsplex from Rat Liver Mitochondria, Arch. Biochem. Biophys., 1972, vol. 149, pp. 425–434.

    Article  PubMed  Google Scholar 

  • Van Tuyle, G.C. and McPherson, M.L., A Compact Form of Rat Liver Mitochondrial DNA Stabilized by Bound Proteins, J. Biol. Chem., 1979, vol. 254, pp. 6044–6053.

    PubMed  Google Scholar 

  • Wallace, D.C., Diseases of the Mitochondrial DNA, Annu. Rev. Biochem., 1992, vol. 61, pp. 1175–1212.

    Article  PubMed  Google Scholar 

  • Wang, N., Gottesman, S., Willingham, M.C., Gottesman, M., and Maurizi, M., A Human Mitochondrial ATP-Dependent Protease That Is Highly Homologous To Bacterial Lon Protease, Proc. Natl. Acad. Sci. U.S.A., 1993, vol. 90, pp. 11247–11251.

    PubMed  Google Scholar 

  • Yamada, E.W., Dotzlaw, H., and Huzel, N.J., Isolation of Histone-Like Proteins from Mitochondria of Bovine Heart, Prepar. Biochem., 1991, vol. 21, pp. 11–23.

    Google Scholar 

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Authors and Affiliations

  1. Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Pushchino, Moscow oblast, 142290, Russia

    M. P. Kutsyi, N. A. Gouliaeva, E. A. Kuznetsova & A. I. Gaziev

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  1. M. P. Kutsyi
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  2. N. A. Gouliaeva
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  3. E. A. Kuznetsova
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  4. A. I. Gaziev
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Translated from Izvestiya Akademii Nauk, Seriya Biologicheskaya, No. 5, 2005, pp. 531–538.

Original Russian Text Copyright © 2005 by Kutsyi, Gouliaeva, Kuznetsova, Gaziev.

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Kutsyi, M.P., Gouliaeva, N.A., Kuznetsova, E.A. et al. Study of DNA-Binding Proteins of Rat Liver Mitochondria. Biol Bull Russ Acad Sci 32, 438–444 (2005). https://doi.org/10.1007/s10525-005-0121-3

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