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RNA
2010 May 01;165:1078-85. doi: 10.1261/rna.2060210.
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Regulation of pre-mRNA splicing in Xenopus oocytes by targeted 2'-O-methylation.
Ge J
,
Liu H
,
Yu YT
.
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The 2'-OH group of the branch point adenosine is a key moiety to initiate pre-mRNA splicing. We use RNA-guided RNA modification to target the pre-mRNA branch point adenosine for 2'-O-methylation, with the aim of blocking pre-mRNA splicing in vertebrate cells. We show that, under certain conditions, injection of a branch point-specific artificial box C/D RNA into Xenopus oocytes effectively 2'-O-methylates adenovirus pre-mRNA at the target nucleotide. However, 2'-O-methylation at the authentic branch point activates a host of cryptic branch points, thus allowing splicing to continue. These cryptic sites are mapped, and mutated. Upon injection, pre-mRNA free of cryptic branch points fails to splice when the branch point-specific box C/D RNA is present. However, 2'-O-methylation at the branch point does not prevent pre-mRNA from being assembled into pre-catalytic spliceosome-like complexes prior to the first chemical step of splicing. Our results demonstrate that RNA-guided pre-mRNA modification can occur in the nucleoplasm of vertebrate cells, thus offering a powerful tool for molecular biology research.
Altafaj,
Oocyte expression with injection of purified T7 RNA polymerase.
2006, Pubmed,
Xenbase
Altafaj,
Oocyte expression with injection of purified T7 RNA polymerase.
2006,
Pubmed
,
Xenbase
Cavaillé,
Targeted ribose methylation of RNA in vivo directed by tailored antisense RNA guides.
1996,
Pubmed
Darzacq,
Cajal body-specific small nuclear RNAs: a novel class of 2'-O-methylation and pseudouridylation guide RNAs.
2002,
Pubmed
,
Xenbase
Deryusheva,
Small Cajal body-specific RNAs of Drosophila function in the absence of Cajal bodies.
2009,
Pubmed
,
Xenbase
Kiss,
Small nucleolar RNAs: an abundant group of noncoding RNAs with diverse cellular functions.
2002,
Pubmed
Kiss-László,
Site-specific ribose methylation of preribosomal RNA: a novel function for small nucleolar RNAs.
1996,
Pubmed
Konarska,
Analysis of splicing complexes and small nuclear ribonucleoprotein particles by native gel electrophoresis.
1989,
Pubmed
Liang,
The spliced leader-associated RNA is a trypanosome-specific sn(o) RNA that has the potential to guide pseudouridine formation on the SL RNA.
2002,
Pubmed
Liu,
Probing RNA in vivo with methylation guide small nucleolar RNAs.
2001,
Pubmed
Liu,
Interference probing of rRNA with snoRNPs: a novel approach for functional mapping of RNA in vivo.
2004,
Pubmed
Liu,
Mis-targeted methylation in rRNA can severely impair ribosome synthesis and activity.
2008,
Pubmed
Newby,
Sculpting of the spliceosomal branch site recognition motif by a conserved pseudouridine.
2002,
Pubmed
Ni,
SnoRNAs as tools for RNA cleavage and modification.
1997,
Pubmed
Nilsen,
RNA-RNA interactions in the spliceosome: unraveling the ties that bind.
1994,
Pubmed
Parker,
Recognition of the TACTAAC box during mRNA splicing in yeast involves base pairing to the U2-like snRNA.
1987,
Pubmed
Query,
Branch nucleophile selection in pre-mRNA splicing: evidence for the bulged duplex model.
1994,
Pubmed
Ruskin,
An RNA processing activity that debranches RNA lariats.
1985,
Pubmed
Ruskin,
Excision of an intact intron as a novel lariat structure during pre-mRNA splicing in vitro.
1984,
Pubmed
Ruskin,
RNA lariat debranching enzyme as tool for analyzing RNA structure.
1990,
Pubmed
Semenov,
Splicing by exon exclusion impaired by artificial box c/d RNA targeted to branch-point adenosine.
2008,
Pubmed
Smith,
Sno storm in the nucleolus: new roles for myriad small RNPs.
1997,
Pubmed
Staley,
Mechanical devices of the spliceosome: motors, clocks, springs, and things.
1998,
Pubmed
Tycowski,
A small nucleolar RNA requirement for site-specific ribose methylation of rRNA in Xenopus.
1996,
Pubmed
,
Xenbase
Tycowski,
A conserved WD40 protein binds the Cajal body localization signal of scaRNP particles.
2009,
Pubmed
Wu,
Mammalian pre-mRNA branch site selection by U2 snRNP involves base pairing.
1989,
Pubmed
Yu,
Internal modification of U2 small nuclear (sn)RNA occurs in nucleoli of Xenopus oocytes.
2001,
Pubmed
,
Xenbase
Yu,
Modifications of U2 snRNA are required for snRNP assembly and pre-mRNA splicing.
1998,
Pubmed
,
Xenbase
Zhao,
An H/ACA guide RNA directs U2 pseudouridylation at two different sites in the branchpoint recognition region in Xenopus oocytes.
2002,
Pubmed
,
Xenbase
Zhao,
Pseudouridines in and near the branch site recognition region of U2 snRNA are required for snRNP biogenesis and pre-mRNA splicing in Xenopus oocytes.
2004,
Pubmed
,
Xenbase
Zhao,
Targeted pre-mRNA modification for gene silencing and regulation.
2008,
Pubmed
Zhuang,
A compensatory base change in human U2 snRNA can suppress a branch site mutation.
1989,
Pubmed
