Biochemical, molecular genetic and high resolution X-ray crystallographic experiments will be used to establish the structural bases for the functional mechanisms by which the ribosome and associated factors achieve their roles in protein synthesis and aminoacyl-tRNA synthetases attach .the correct amino acid to the right tRNA. To further illuminate the mechanisms of each step in the process of protein synthesis, we wish to obtain crystals of the ribosome trapped in as many of the steps in the protein synthesis cycle as possible and to establish their structures at the highest resolution possible. The complexes whose structures will be pursued include those of the 1) 70S ribosome with bound m-RNA, tRNA and either elongation factor Tu with aminoacyl-tRNA or elongation factor G, 2) complexes between the 70S ribosomal and a P-site tRNA containing a translation arresting polypeptide, and 3) complexes between the 70S ribosome the protein secretion and membrane protein insertion channel, the translocon. To explore the differences between the bacterial ribosome and its much large counterpart from eukaryotes, particularly in the initiation step of protein synthesis, the structures of the 40S subunit, the 60S subunit and/or the SOS ribosome, including an initiation complex formed with IRIS RNA, and using ribosomes isolated from either yeast or rabbit reticulocyte will be pursued. In order to understand how aminoacyl-tRNA synthetases, such as Gln-tRNA synthetase, are stimulated to aminoacylate the tRNA upon-anticodon recognition, structures of complexes with tRNAs containing mutated anticodons will be established.
A mechanistic understanding of the process of translation from the specific charging of tRNA to all aspects of protein synthesis is fundamental in biology. Both aminoacyl-tRNA synthetases and the ribosome are major targets of clinically relevant antibacterial drugs, whose further novel development will be facilitated by these studies.
|Meehan, Robert E; Torgerson, Chad D; Gaffney, Barbara L et al. (2016) Nuclease-Resistant c-di-AMP Derivatives That Differentially Recognize RNA and Protein Receptors. Biochemistry 55:837-49|
|Li, Sanshu; Hwang, Xue Ying; Stav, Shira et al. (2016) The yjdF riboswitch candidate regulates gene expression by binding diverse azaaromatic compounds. RNA 22:530-41|
|Liu, Bin; Zuo, Yuhong; Steitz, Thomas A (2016) Structures of E. coli ÏƒS-transcription initiation complexes provide new insights into polymerase mechanism. Proc Natl Acad Sci U S A 113:4051-6|
|Askerka, Mikhail; Wang, Jimin; Vinyard, David J et al. (2016) S3 State of the O2-Evolving Complex of Photosystem II: Insights from QM/MM, EXAFS, and Femtosecond X-ray Diffraction. Biochemistry 55:981-4|
|Gagnon, Matthieu G; Lin, Jinzhong; Steitz, Thomas A (2016) Elongation factor 4 remodels the A-site tRNA on the ribosome. Proc Natl Acad Sci U S A 113:4994-9|
|Ruff, Karen M; Muhammad, Ayesha; McCown, Phillip J et al. (2016) Singlet glycine riboswitches bind ligand as well as tandem riboswitches. RNA 22:1728-1738|
|Furukawa, Kazuhiro; Ramesh, Arati; Zhou, Zhiyuan et al. (2015) Bacterial riboswitches cooperatively bind Ni(2+) or Co(2+) ions and control expression of heavy metal transporters. Mol Cell 57:1088-98|
|Roy, Raktim N; Lomakin, Ivan B; Gagnon, Matthieu G et al. (2015) The mechanism of inhibition of protein synthesis by the proline-rich peptide oncocin. Nat Struct Mol Biol 22:466-9|
|Polikanov, Yury S; Moore, Peter B (2015) Acoustic vibrations contribute to the diffuse scatter produced by ribosome crystals. Acta Crystallogr D Biol Crystallogr 71:2021-31|
|Weinberg, Zasha; Kim, Peter B; Chen, Tony H et al. (2015) New classes of self-cleaving ribozymes revealed by comparative genomics analysis. Nat Chem Biol 11:606-10|
Showing the most recent 10 out of 100 publications