Abstract
The crystal structure of mitochondrial F1-ATPase indicatesthat the α and β subunits fold into a structure defined by threedomains: the top β-barrel domain, the middle nucleotide-binding domain,and the C-terminal α-helix bundle domain (Abraham et al.1994); Bianchet et al., 1998). The β-barrel domains of theα and β subunits form a crown structure at the top ofF1, which was suggested to stabilize it (Abraham et al.1994). In this study. the role of the β-barrel domain in the α andβ subunits of the yeast Saccharomyces cerevisiae F1,with regard to its folding and assembly, was investigated. The β-barreldomains of yeast F1 α and β subunits were expressedindividually and together in Escherichia coli. When expressedseperately, the β-barrel domain of the β subunit formed a largeaggregate structure, while the domain of the α subunit waspredominately a monomer or dimer. However, coexpression of the β-barreldomain of α subunit domain. Furthermore, the two domains copurified incomplexes with the major portion of the complex found in a small molecularweight form. These results indicate that the β-barrel domain of theα and β subunits interact specifically with each other and thatthese interactions prevent the aggregation of the β-barrel domain of theβ subunit. These results mimic in vivo results and suggest thatthe interactions of the β-barrel domains may be critical during thefolding and assembly of F1.
REFERENCES
Abrahams, J. P., Leslie, A. G. W., Lutter, R., and Walker, J. E. (1994). Nature 370, 621–628.
Ackerman, S. H. and Tzagoloff, A. (1990). Proc. Natl. Acad. Sci. USA 87, 4986–4990.
Ackerman, S. H., Martin, J., and Tzagoloff, A. (1992). J. Biol. Chem. 267, 7386–7394.
Avni A., Avital S., and Gromet-Elhanan Z. (1991). J. Biol. Chem. 266, 7317–7320.
Bianchet, M. A., Hullihen, J., Pedersen, P. L., and Amzel, L. M. (1998). Proc. Natl. Acad. Sci. USA 95, 11065–11070, 1998.
Blackwell, T. K., Bowerman, B., Priess, J. R., and Weintraub, H. (1994). Science 266, 621–628.
Brahms, S. and Brahms, J. (1980). J. Mol. Biol. 138, 149–178.
Cheng, M., Hartl, F.-U., Martin, J., Pollock, R., Kalousek, F., Neupert, W., and Horowich, A. (1989). Nature 337, 620–625.
Chiang, C. M. and Roeder, R. G. Peptide Res. 6, 62–64.
Frankel, A. D. and Smith, C. A. (1998). Cell 92, 149–151.
Genetics Computer Group (1996). Wisconsin Package, Version 8, Madison, WI.
Gao F., Lipscomb, B., Wu, I., and Richter, M. L. (1995). J. Biol. Chem 270, 9763–9769.
Kang, P. J., Ostermann, J., Shilling, J., Neupert, W., Craig, E. A., and Pfanner, N. (1990). Nature 348, 137–143.
Laemmli, U. K. (1970). Nature 227, 680–685.
Maniatis, T., Fritsch, D. F., and Sambrook, J. (1982). In Molecular Cloning, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York), pp. 368–369.
Mukhopadhyay, A., Zhou, X., Uh, M., and Mueller, D. M. (1992). J. Biol. Chem. 267, 25690–25696.
Osterman, J., Horowich, A. L., Neupert, W., and Hartly F. U. (1989). Nature 341, 125–130.
Perczel, A., Park, K., and Fasman, G. D. (1992). Anal. Biochem. 203, 83–93.
Sanger, F., Nicklen, S., and Coulson, A. R. (1977). Proc. Natl. Acad. Sci. USA 74, 5463–5467.
Schagger, H. and Von Jagow, G. (1987). Anal. Biochem. 166, 368–379.
Spolar, R. S. and Record, M. T. (1994). Science 263, 777–784.
Stoscheck, C. M. (1990). Methods Enzymol. 182, 50–68.
Studier, F. W., Rosenberg, A. H., Dunn, J. J., and Dubendorff, J. W. (1990). Methods Enzymol. 185, 61–89.
Towbin, H., Staehelin, T., and Gordin, J. (1979). Proc. Natl. Acad. Sci. USA 76, 4350–4354.
Vogel, G. and Steinhart R., (1976). Biochemistry 15, 208–216.
Walker, J. E., Fearnley, I. M., Gay, N. J., Gibson, B. W., Northrop, F. D., Powell, S. J., Runswick, M. J., Saraste, M., and Tybulewicz, V. L. J. (1985). J. Mol. Biol. 184, 677–701.
Yoshida, M., Okamoto, H., Sone, N., Hirata, H., and Kagawa, Y. (1977). Proc. Natl. Acad. Sci. USA 74, 936–940.
Yuan, H. and Douglas, M. G. (1992). J. Biol. Chem. 267, 14697–14702.
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Yao, B., Mueller, D.M. The Role of the Amino-Terminal β-Barrel Domain of the α and β Subunits in the Yeast F1-ATPase. J Bioenerg Biomembr 31, 95–104 (1999). https://doi.org/10.1023/A:1005443609985
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DOI: https://doi.org/10.1023/A:1005443609985