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1. Introduction
2. Talk outline
3. Modified nucleotides in RNA (1)
4. Modified nucleotides in RNA (2)
5. The ribosome components
6. Distribution of modified nucleotides in the ribosome
7. Incorporation of modified nucleotides into ribosome
8. Enzymatic synthesis of modified RNAs
9. Chemical synthesis of modified RNAs
10. Protective group strategies
11. Phosphoramidites
12. Solid support
13. Stages of chemical synthesis of modified RNAs
14. Novel protective groups on target phosphoramidites
15. Chemical synthesis of RNA with ACE/BzH
16. Deprotection of synthetic RNAs
17. Why chemical RNA synthesis?
18. Semi-synthesis of modified RNAs
19. Modified sites in the 3' half of E. coli 23S rRNA
20. The 1920 region (H69) of E. coli 23S rRNA
21. Interface region between ribosomal subunits
22. Thermus thermophilus 70S ribosome
23. Highly conserved modification sites in H70
24. Modified sites in E. coli 16S rRNA
25. 2'-O-ACE-5'-O-BzH-protected phosphoramidites
26. 15N-Labeled pseudouridine derivatives
27. 2'-O-ACE-5'-O-BzH-pseudouridine phosphoramidite
28. 3-methyl-2'-O-ACE-5'-O-BzH-pseudouridine
29. [3-15N]-3-methylpseudouridine derivative synthesis
30. 5'-O-BzH-2'-O-ACE- [3-15N,1-15N] synthesis
31. Synthesis of m1acp3-Psi
32. Long-term goals of our research program
33. RNA hairpins representing the 1920 loop (H69)
34. Determining the stability of the H69 RNAs
35. H69 RNAs with Psi modifications thermodynamics
36. Psi, m3U, or m3 Psi modifications (1)
37. Psi, m3U, or m3 Psi modifications (2)
38. Effects of pH and sequence on H69 stability
39. Typical chemical shifts in NMR spectroscopy
40. 1D imino proton NMR spectra at 3 degrees C (1)
41. 1D imino proton NMR spectra at 3 degrees C (2)
42. Synthesis of 15N-labeled RNAs
43. 15N-H splitting
44. NMR spectrum of (15N)U(15N)U(15N)U
45. Imino proton spectra of (15N)-Psi-Psi-Psi at 3 degrees C
46. Imino proton spectra of Psi-(15N)-Psi-Psi at 3 degrees C
47. Imino proton spectra of Psi-Psi-(15N)-Psi at 3 degrees C
48. NMR results with 15N-labeled RNAs
49. 1D imino proton NMR spectra of human analogues
50. Conclusions
51. Acknowledgements

Topics Covered

• Background on modified RNAs
• Approaches for generating site-specifically modified RNAs
• Examples of modifications (pseudouridine) in the ribosome
• Biophysical studies on site-specifically modified RNAs representing helix 69 of the ribosome

Links

Series:

• Chemical Biology

Categories:

• Biochemistry

Talk Citation

Publication History

• Published on August 14, 2008
• Reviewed on June 30, 2021

Financial Disclosures

• Prof. Christine Chow has not informed HSTalks of any commercial/financial relationship that it is appropriate to disclose.