Gen511-3-10-08F

Gen511-3-10-08F - Gen511: Monday 10 March 2008 Chapter 14:...

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Gen511: Monday 10 March 2008 Chapter 14: Splicing Genetic analysis of U2-branchpoint interaction Association of snRNPs with pre-mRNA Splicing protein interactions: a. Synthetic lethal screens b. yeast 2-hybrid Role of RNA PolII CTD How are multiple introns spliced? Alternative splicing
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14-2 Mechanism of intron excision
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Splicing is Essential: most eukaryotic genes contain introns that interrupt the ORF. Splicing is required to restore the ORF and generate functional proteins. Hence, splicing is difficult to study genetically (mutants are lethal). Yeast: can isolate conditional lethals (e.g. temperature sensitive) that survive under permissive conditions (e.g. 30 degrees) but die in restrictive conditions (e.g. 37 degrees). Example: prp mutants (pre-RNA processing) accumulate unspliced precursor RNA. Some of these prp mutants have mutations in proteins that make up the spliceosome. Genetic analysis of splicing
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14-4 How do snRNPs promote splicing? They help to position three reacting sites in precursor mRNA: 5’ splice donor site Branch point sequence 3’ splice acceptor site Base-pairing between snRNAs and the pre- mRNA sequences are critical.
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Review of snRNA associations U1 base pairs with the 5’ splice site U2 base pairs with the conserved sequence at the splicing branchpoint U4 base pairs with U6 U6 base pairs with the 5’ splice site, and with U2 U5 associates with the last nucleotide in one exon and the first nucleotide of the next, presumably lining up the exons for splicing
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U2 snRNA base- pairs with the conserved sequence at the splicing branchpoint. This base pairing is essential for splicing. What’s the evidence
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This note was uploaded on 03/31/2008 for the course GEN 511 taught by Professor Peterson during the Spring '08 term at Iowa State.

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Gen511-3-10-08F - Gen511: Monday 10 March 2008 Chapter 14:...

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