Blaha Gregor, Dr. techn. post doctoral fellow Center for Structural Biology Department of Molecular Biophysics & Biochemistry Yale University 266 Whitney Avenue, 415 Bass Center New Haven, CT 06520-8114 Phone:  (203) 432-5626 FAX:  (203) 432-3282 Email:   gregor.blaha{at}yale.edu

Summary:

The ribosome plays a central role in the translation of the genetic information into proteins. The bacterial 70S ribosome consists of the smaller 30S and larger 50S subunit. The structure of both subunits was solved to atomic resolution (1, 2, 3, 4). A 5.5 Å resolution structure of 70S in complex with tRNA and mRNA was published. However, at this resolution only the backbone of the rRNA and the ribosomal proteins could be traced (5). So far the best resolution for a ribosomal subunits or its complexes is from the work with 50S from H. marismortui (1).
In an effort to improve the resolution of the ribosomal structure, we set out to systematically investigate the conditions of in vitro complex formation with ribosomes from extremophiles. We have tried to obtain a homogenous population of 70S complexes with sucrose gradient centrifugations and hydrophobic interaction chromatography. Further experiments are being performed to increase the homogeneity of preparations and to establish crystallization conditions for those preparations.

1. Ban, N., Nissen, P., Hansen, J., Moore, P.B., Steitz, T.A. (2000) The complete atomic structure of the large ribosomal subunit at 2.4 A resolution. Science 289,905-20.
2. Wimberly, B.T., Brodersen, D.E., Clemons, W.M. Jr., Morgan-Warren, R.J., Carter, A.P., Vonrhein, C., Hartsch, T. and Ramakrishnan, V. (2000) Structure of the 30S ribosomal subunit. Nature 407, 327-39.
3. Schluenzen, F., Tocilj, A., Zarivach, R., Harms, J., Gluehmann, M., Janell, D., Bashan, A., Bartels, H., Agmon, I., Franceschi, F. and Yonath A. (2000) Structure of functionally activated small ribosomal subunit at 3.3 angstroms resolution. Cell 102, 615-23.
4. Harms, J., Schluenzen, F., Zarivach, R., Bashan, A., Gat, S., Agmon, I., Bartels, H., Franceschi, F. and Yonath, A. (2001) High resolution structure of the large ribosomal subunit from a mesophilic eubacterium. Cell 107, 679-88.
5. Yusupov, M.M., Yusupova, G.Z., Baucom, A., Lieberman, K., Earnest, T.N., Cate, J.H. and Noller, H.F. (2001) Crystal structure of the ribosome at 5.5 A resolution. Science 292, 883-96.

Publication:

Blaha G., Wilson D.N., Stoller G., Fischer G., Willumeit R., Nierhaus KH: "Localization of the Trigger Factor Binding Site on the Ribosomal 50S Subunit" (in press)