Structure and structural variations of the Escherichia coli 30 S ribosomal subunit as revealed by three-dimensional cryo-electron microscopy.

A three-dimensional reconstruction of the 30 S subunit of the Escherichia coli ribosome was obtained at 23 A resolution. Because of the improved resolution, many more structural details are seen as compared to those obtained in earlier studies. Thus, the new structure is more suitable for comparison with the 30 S subunit part of the 70 S ribosome, whose structure is already known at a better resolution. In addition, we observe relative and, to some extent, independent movements of three main structural domains of the 30 S subunit, namely head, platform and the main body, which lead to partial blurring of the reconstructed volume. An attempt to subdivide the data set into conformationally defined subsets reveals the existence of conformers in which these domains have different orientations with respect to one another. This result suggests the existence of dynamic properties of the 30 S subunit that might be required for facilitating its interactions with mRNA, tRNA and other ligands during protein biosynthesis.

[1]  C. Spahn,et al.  Protection Patterns of tRNAs Do Not Change during Ribosomal Translocation* , 1998, The Journal of Biological Chemistry.

[2]  J Frank,et al.  Escherichia coli 70 S ribosome at 15 A resolution by cryo-electron microscopy: localization of fMet-tRNAfMet and fitting of L1 protein. , 1998, Journal of molecular biology.

[3]  Joachim Frank,et al.  A 9 Å Resolution X-Ray Crystallographic Map of the Large Ribosomal Subunit , 1998, Cell.

[4]  J Frank,et al.  Visualization of elongation factor G on the Escherichia coli 70S ribosome: the mechanism of translocation. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[5]  S. Mottagui-Tabar,et al.  Evidence for in vivo ribosome recycling, the fourth step in protein biosynthesis , 1998, The EMBO journal.

[6]  A. Spirin,et al.  In vitro assembly of a ribonucleoprotein particle corresponding to the platform domain of the 30S ribosomal subunit. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[7]  P. Hagerman,et al.  Protein and Mg(2+)-induced conformational changes in the S15 binding site of 16 S ribosomal RNA. , 1998, Journal of molecular biology.

[8]  R. Brimacombe,et al.  Visualization of elongation factor Tu on the Escherichia coli ribosome , 1997, Nature.

[9]  R. Brimacombe,et al.  A new model for the three-dimensional folding of Escherichia coli 16 S ribosomal RNA. II. The RNA-protein interaction data. , 1997, Journal of molecular biology.

[10]  M van Heel,et al.  A new model for the three-dimensional folding of Escherichia coli 16 S ribosomal RNA. III. The topography of the functional centre. , 1997, Journal of molecular biology.

[11]  A E Dahlberg,et al.  A conformational switch in Escherichia coli 16S ribosomal RNA during decoding of messenger RNA. , 1997, Science.

[12]  R. Brimacombe,et al.  A new model for the three-dimensional folding of Escherichia coli 16 S ribosomal RNA. I. Fitting the RNA to a 3D electron microscopic map at 20 A. , 1997, Journal of molecular biology.

[13]  D. Draper,et al.  Effects of Mg2+, K+, and H+ on an equilibrium between alternative conformations of an RNA pseudoknot. , 1997, Journal of molecular biology.

[14]  R. Brimacombe,et al.  The path of mRNA through the bacterial ribosome: a site-directed crosslinking study using new photoreactive derivatives of guanosine and uridine. , 1997, RNA.

[15]  J. Frank,et al.  Three-dimensional reconstruction with contrast transfer function correction from energy-filtered cryoelectron micrographs: procedure and application to the 70S Escherichia coli ribosome. , 1997, Journal of structural biology.

[16]  R. Brimacombe,et al.  Arrangement of tRNAs in Pre- and Posttranslocational Ribosomes Revealed by Electron Cryomicroscopy , 1997, Cell.

[17]  J Frank,et al.  Three-dimensional reconstruction of the Escherichia coli 30 S ribosomal subunit in ice. , 1996, Journal of molecular biology.

[18]  R. Agrawal,et al.  Sites of Ribosomal RNAs Involved in the Subunit Association of Tight and Loose Couple Ribosomes* , 1996, The Journal of Biological Chemistry.

[19]  C. Gualerzi,et al.  Late events in translation initiation. Adjustment of fMet-tRNA in the ribosomal P-site. , 1996, Journal of molecular biology.

[20]  J. Frank,et al.  Direct Visualization of A-, P-, and E-Site Transfer RNAs in the Escherichia coli Ribosome , 1996, Science.

[21]  R. Brimacombe,et al.  From stand-by to decoding site. Adjustment of the mRNA on the 30S ribosomal subunit under the influence of the initiation factors. , 1995, RNA.

[22]  D. Draper,et al.  Messenger RNA Recognition by Fragments of Ribosomal Protein S4 (*) , 1995, The Journal of Biological Chemistry.

[23]  J. Frank,et al.  A model of protein synthesis based on cryo-electron microscopy of the E. coli ribosome , 1995, Nature.

[24]  M van Heel,et al.  The 70S Escherichia coli ribosome at 23 A resolution: fitting the ribosomal RNA. , 1995, Structure.

[25]  H. Noller,et al.  Independent in vitro assembly of a ribonucleoprotein particle containing the 3' domain of 16S rRNA. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[26]  S C Harvey,et al.  A quantitative model of the Escherichia coli 16 S RNA in the 30 S ribosomal subunit. , 1994, Journal of molecular biology.

[27]  M. Laughrea Structural dynamics of translating ribosomes: 16S ribosomai RNA bases that may move twice during translocation , 1994, Molecular microbiology.

[28]  J. Frank,et al.  The ribosome at improved resolution: new techniques for merging and orientation refinement in 3D cryo-electron microscopy of biological particles. , 1994, Ultramicroscopy.

[29]  R. Albrecht-Ehrlich,et al.  Immunoelectron microscopic localization of ribosomal proteins BS8, BS9, BS20, BL3 and BL21 on the surface of 30S and 50S subunits from Bacillus stearothermophilus. , 1993, European journal of biochemistry.

[30]  J. Ofengand,et al.  Chemical evidence for domain assembly of the Escherichia coli 30S ribosome , 1993, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[31]  D. P. Burma,et al.  Mechanism of ribosomal subunit association: oligodeoxynucleotides as probes. , 1992, Indian journal of biochemistry & biophysics.

[32]  A. Spirin,et al.  Structural dynamics of translating ribosomes. , 1992, Biochimie.

[33]  W. Hill,et al.  Probing dynamic changes in rRNA conformation in the 30S subunit of the Escherichia coli ribosome. , 1992, Biochemistry.

[34]  C. Gualerzi,et al.  Subunit association defects in Escherichia coli ribosome mutants lacking proteins S20 and L11. , 1989, European journal of biochemistry.

[35]  P. Wollenzien,et al.  An RNA secondary structure switch between the inactive and active conformations of the Escherichia coli 30 S ribosomal subunit. , 1989, The Journal of biological chemistry.

[36]  H. Noller,et al.  Model for the three-dimensional folding of 16 S ribosomal RNA. , 1988, Journal of molecular biology.

[37]  R. Brimacombe,et al.  A detailed model of the three-dimensional structure of Escherichia coli 16 S ribosomal RNA in situ in the 30 S subunit. , 1988, Journal of molecular biology.

[38]  J A Langer,et al.  A complete mapping of the proteins in the small ribosomal subunit of Escherichia coli. , 1987, Science.

[39]  J. Bachellerie,et al.  Higher-order structure of domain III in Escherichia coli 16S ribosomal RNA, 30S subunit and 70S ribosome. , 1987, Biochimie.

[40]  K. Wong,et al.  The role of magnesium and potassium ions in the molecular mechanism of ribosome assembly: hydrodynamic, conformational, and thermal stability studies of 16 S RNA from Escherichia coli ribosomes. , 1986, Archives of biochemistry and biophysics.

[41]  T. Ohta,et al.  Fluorometric investigation of the local conformation of 16S rRNA in the regions of the 3'- and 5'-ends. , 1976, Journal of biochemistry.

[42]  J. Frank,et al.  Conformational variability in Escherichia coli 70S ribosome as revealed by 3D cryo-electron microscopy. , 1999, The international journal of biochemistry & cell biology.

[43]  R. Jackson,et al.  A prokaryotic-like mode of cytoplasmic eukaryotic ribosome binding to the initiation codon during internal translation initiation of hepatitis C and classical swine fever virus RNAs. , 1998, Genes & development.

[44]  Walter E. Hill,et al.  The Ribosome : structure, function, and evolution , 1990 .

[45]  D. Camp,et al.  Probing ribosome structure and function using short oligodeoxyribonucleotides. , 1988, Methods in enzymology.

[46]  L. Gorini Streptomycin and Misreading of the Genetic Code , 1974 .