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1. Introduction
2. RNAi - type processes
3. Creation of siRNA and miRNA
4. Initiation and effector stage
5. Another pathway of gene silencing by RISC
6. Key proteins in RNAi - Dicer and argonaute
7. Two subclasses of Ago proteins
8. The Argonuate protein family - PAZ
9. Crystal and NMR structures of PAZ domains
10. Ago2-PAZ
11. The PAZ domain has a deviant OB-fold
12. Ago2-PAZ - second sub-domain
13. siRNA contact Argonaute proteins in RISC
14. Ago2-PAZ binds 3' ends and 3' overhangs
15. Invariant residues in the PAZ domain
16. Aromatic residues involved in RNA binding
17. PAZ recognizes the 3'-end of an siRNA
18. PAZ 3'-end recognition
19. The Argonaute protein family - PIWI
20. Structure of Argonaute from Pyrococcus furiousus
21. Rotation of the Argonaute structure
22. The PAZ domain of Argonaute
23. PAZ domains of pf-Ago and hAgo1
24. Sequences of PAZ domains of pf-Ago and hAgo1
25. Structure of Argonaute from Pyrococcus furiousus
26. The PIWI domain
27. PIWI is an RNase H domain
28. PIWI and RNase H have similar active sites
29. DDE motif is conserved (1)
30. RISC vs. RNase H
31. How the RNA would bind to Argonaute? (1)
32. How the RNA would bind to Argonaute? (2)
33. A binding groove for RNA?
34. RNA falls in the large groove
35. A model for mRNA slicing
36. Substrate loading and release (1)
37. Substrate loading and release (2)
38. Argonaute as slicer
39. Only Ago2 makes a cealving RISC
40. DDE motif is conserved (2)
41. Inactivating slicer activity
42. The active site residues - DDE motif?
43. Argonaute has a novel "DDH" motif
44. The "DDH" motif in PIWI
45. Ago2 and an siRNA form recombinant RISC
46. Recombinant RISC is a Mg dependant enzyme
47. 5'-phosphate stabilizes recombinant RISC
48. Cleavage position is measured from siRNA's 5'-end
49. Cleavage by recombinant RISC is accurate
50. dsRNA binding to Af-PIWI domain protein (1)
51. A model for 5'-phosphate binding
52. dsRNA binding to Af-PIWI domain protein (2)
53. Kinetics
54. ATP-dependant product release by RISC
55. ATP does not accelerate cleavage by RISC
56. Summary

Topics Covered

• Impact of RNAi on biology
• Gene silencing
• Mechanism of unravelling
• Understanding the mechanism of RNAi related pathways
• Transcriptional and post-transcriptional gene silencing
• DNA elimination in tetrahymena
• Mechanistic aspects of RNAi
• The argonaute protein

Links

Series:

• RNA Interference

Categories:

• Biochemistry
• Cell Biology

Talk Citation

Joshua-Tor, L. (2020, September 10). Mechanisms of RNAi: a molecular view or Argonaute: the secret life of Slicer [Video file]. In The Biomedical & Life Sciences Collection, Henry Stewart Talks. Retrieved April 16, 2024, from https://hstalks.com/bs/643/.
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Publication History

• Published on October 1, 2007
• Reviewed on September 10, 2020

Financial Disclosures

• Prof. Leemor Joshua-Tor has not informed HSTalks of any commercial/financial relationship that it is appropriate to disclose.