2008
DOI: 10.1261/rna.1189008
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Mutational analysis of the U12-dependent branch site consensus sequence

Abstract: Highly conserved sequences at the 59 splice site and branch site of U12-dependent introns are important determinants for splicing by U12-dependent spliceosomes. This study investigates the in vivo splicing phenotypes of mutations in the branch site consensus sequence of the U12-dependent intron F from a human NOL1 (P120) minigene. Intron F contains a fully consensus branch site sequence (UUCCUUAAC). Mutations at each position were analyzed for their effects on U12-dependent splicing in vivo. Mutations at most … Show more

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Cited by 8 publications
(16 citation statements)
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“…The U11 GG6/7CC, U6atac GG14/15CC and U12 GA23/24CU expression plasmids have been described previously 10 11 12 14 23 24 36 . Second site mutations were introduced in U6atac GG14/15CC and U12 GA23/24CU snRNAs to construct swapped SL snRNA plasmids and U6atac distal 3′ SL mutant plasmids.…”
Section: Methodsmentioning
confidence: 99%
“…The U11 GG6/7CC, U6atac GG14/15CC and U12 GA23/24CU expression plasmids have been described previously 10 11 12 14 23 24 36 . Second site mutations were introduced in U6atac GG14/15CC and U12 GA23/24CU snRNAs to construct swapped SL snRNA plasmids and U6atac distal 3′ SL mutant plasmids.…”
Section: Methodsmentioning
confidence: 99%
“…The starting U12 snRNA expression plasmid has been described previously ( 8 ). This construct contains mutations GA23/24CU in the branch site interacting region to restore complementary base pairing to UC84/85AG branch site region of P120 U12-dependent intron F. For this study, second site mutations were introduced in U12 snRNA in the background of GA23/24CU.…”
Section: Methodsmentioning
confidence: 99%
“…U2 snRNA binds the branch site sequences by canonical base pairing interactions, which are necessary for U2-dependent intron splicing ( 7 ). Similarly, U12 snRNA binds to the branch site of a U12-dependent intron by RNA-RNA base pairing essential for splicing ( 3 , 8 ). Besides binding to the branch site sequence, U2 snRNA also base pairs with U6 snRNA to form an intermolecular helix required for U2-dependent splicing ( 9–12 ).…”
Section: Introductionmentioning
confidence: 99%
“…Naturally occurring U12‐type introns mostly belong to either the AT–AC or GT–AG subtypes. A few natural examples of U12‐type introns with other combinations of terminal residues have been reported, and non‐canonical termini have also been shown to support U12‐dependent splicing experimentally, although often with reduced efficiency 11–15. However, combinations of the main subtypes (i.e., AT–AG or GT–AC) appear to be disfavored.…”
Section: U12‐type Intron Sequences: Definition and Characteristicsmentioning
confidence: 99%