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Chapter 13

Chapter 13. RNA Splicing (and some other important stuff). 13 and 15 October, 2004. Overview. Introns are removed after transcription - exons may comprise only a few percent of the primary transcript. The location of splicing is determined by splice site consensus sequences.

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Chapter 13

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  1. Chapter 13 RNA Splicing (and some other important stuff) 13 and 15 October, 2004

  2. Overview • Introns are removed after transcription - exons may comprise only a few percent of the primary transcript. • The location of splicing is determined by splice site consensus sequences. • The intron is released as a lariat structure. • Splicing is carried out by a large ribonucleoprotein structure called the spliceosome. • Spliceosome snRNPs use RNA base pairing to recognize splice sites. • Spliceosome components assemble at the 5’ splice site (U1/U6), the Branch point (U2), and the 3’ splice site (U5). • There are at lease two types of self-splicing introns. • Splicing is highly regulated, including many instances of alternative splicing. • Some RNAs are edited before translation. • Polyadenylation is coupled to transport.

  3. Primary transcript and spliced product hybridization suggest splicing.

  4. Splicing

  5. Splicing Consensus Sequences

  6. Unusual Electrophoretic Behavior of Intron and Dependence on snRNPs

  7. The Splicing Reaction

  8. Structure of the Lariat Intermediate

  9. Trans Splicing

  10. snRNP-RNA Recognition

  11. Splicosome Purification

  12. Splicosome Assembly

  13. Base pairing between U2 and branch point sequences is demonstrated by second-site suppressor studies.

  14. 4-thioU crosslinking demonstrates interactions between U6 and the 5’ splice site, U5 and the 3’ splice site.

  15. The Spliceosome Cycle

  16. The two-hybrid system demonstrates interactions between components of the commitment complex.

  17. Self-Splicing Introns

  18. Spliced and Self-splicing Introns

  19. Alternative Splice Sites

  20. Splicing Errors

  21. Regulation of Alternative Splicing

  22. Alternative Splicing

  23. Alternative Splicing

  24. Inhibition of Splicing

  25. AT-AC Spliceosome

  26. Conservation of Splicing Consenses

  27. Exons encode protein domains.

  28. Gene Assembly

  29. Protein Evolution

  30. RNA Editing

  31. C to U deamination

  32. Editing Mechanism

  33. mRNA Transport

  34. Purification of a poly-A binding protein that stimulates poly-A polymerase (PABII)

  35. Model for polyadenylation

  36. Title

  37. Title

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