RNA processing
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Transcript of RNA processing
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RNA processing
Proks vs. EuksIn proks, transcription & translation coupledIn euks, processes are temporally& spatially separated so more control
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RNA processing
mRNA processing in eukaryotes:5’-capping3’-endonuclease cleavage and polyadenylationRNA splicing & editing
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RNA processingmRNA processing:5’-capping
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RNA processingmRNA processing:3’-endonuclease cleavage & polyadenylation
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RNA processing
mRNA processing:RNA splicing & editing
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RNA processingmRNA processing:RNA splicing & editingMainly in eukaryotic organisms, highly conserved from yeast to human
Spliceosome is an RNA-protein complex; RNA proposed to be catalytic
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RNA processing
Alternative splicing
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RNA processing
Alternative splicing
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RNA ProcessingNaturally occurring catalystsCatalytic RNA = Ribozyme
• RNA cleavage
glmS ribozyme - also a RIBOSWITCH!! (crystal structure)hammerhead ribozyme (crystal structure)hairpin ribozyme (crystal structure)Varkud satellite (VS) ribozyme (partial NMR structure)hepatitis delta virus (HDV) ribozyme (crystal structure)M1 RNA (RNase P) (partial crystal structure)
• RNA splicing
group I introns (crystal structure)group II introns (partial crystal structure)*** U2-U6 snRNA (spliceosome) (partial NMR structure) ***
• Peptide bond formation
ribosome (crystal structure)
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RNA processingRNA catalyzed splicing (ribozymes)Group I intron – found in fungi, algae, plants, bacteria
Group I introns found in nuclear, mitochondrial and chloroplast genes of rRNAs, tRNAs and mRNAs
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Group I Intron crystal structure
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RNA processing
RNA catalyzed self-cleavage (ribozymes)Hammerhead ribozyme (found in a virusoids, part of small RNA genome)
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RNA processingRiboswitches
Tom Cech, Nature (2004) 428: 263-264.
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Riboswitches contain an aptamer (RNA binding) domain & expression platform
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Riboswitches
J.K. Soukup and G.A. Soukup, Curr. Opin. Struct. Biol. (2004) 14: 344-349.
Newly discovered riboswitches glycine, pre-Queuosine, 2’-deoxyguanosine, cyclic dimeric GMP
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• Transcription termination and anti-termination• Translation initiation: RBS accessibility• RNA processing: Splicing or degradation
Riboswitch mechanisms
Winkler & Breaker (2005) Annu Rev Microbiol 59: 487-517
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• Recognizes ligand phosphate through metal ion-mediated backbone and nucleobase contacts
• Riboswitch regulation of gene expression is perturbed by the antibiotic PTPP• Riboswitch agonists or antagonists: novel antibiotics?
TPP Riboswitch
Sudarsan et al. (2005) Chem Biol 12:1325-1335; Serganov et al. (2006) Nature 441:1167-1171; Thore et al. (2006) Science 312:1208-1211
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Barrick & Breaker (2006) Scientific American 296: 50-57
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Glucosamine 6-phosphate riboswitch
Klein and Ferré-D'Amaré (2006) Science 313:1752-1756.
Jansen, McCarthy, Soukup & Soukup (2006) Nat Struct Mol Biol 13:517-523.
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A conserved RNA structure precedes the glmS gene in Gram-positive organisms
Kobayashi et al., Proc. Natl. Acad. Sci. USA (2003) 10: 4678-4683.
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RNA Interference (RNAi)
• RNAi is the process whereby double-stranded RNA (dsRNA) induces homology-dependent degradation of cognate RNA (i.e. gene silencing)
• RNAi is highly conserved among eukaryotes (fungi, protozoans, plants, nematodes, invertebrates, mammals)
Biological significance of RNAi
Cellular immune response to viruses
In certain organism (especially plants), RNAi serves as a first line of defense against viral infection, as viral replication typically includes dsRNA species
Genetic stability
RNAi repress the mobility of transposable genetic elements in C. elegans and S. pombe which requires the formation of dsRNA
Organismal development and germline fate
Developmental processes are affected by endogenous non-coding RNAs that function through the RNAi pathway (microRNAs)