Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
RNA
2012 May 01;185:1075-90. doi: 10.1261/rna.031757.111.
Show Gene links
Show Anatomy links
In vitro reconstitution of yeast splicing with U4 snRNA reveals multiple roles for the 3' stem-loop.
Hayduk AJ
,
Stark MR
,
Rader SD
.
???displayArticle.abstract???
U4 small nuclear RNA (snRNA) plays a fundamental role in the process of premessenger RNA splicing, yet many questions remain regarding the location, interactions, and roles of its functional domains. To address some of these questions, we developed the first in vitro reconstitution system for yeast U4 small nuclear ribonucleoproteins (snRNPs). We used this system to examine the functional domains of U4 by measuring reconstitution of splicing, U4/U6 base-pairing, and triple-snRNP formation. In contrast to previous work in human extracts and Xenopus oocytes, we found that the 3' stem-loop of U4 is necessary for efficient base-pairing with U6. In particular, the loop is sensitive to changes in both length and sequence. Intriguingly, a number of mutations that we tested resulted in more stable interactions with U6 than wild-type U4. Nevertheless, each of these mutants was impaired in its ability to support splicing, indicating that these regions of U4 have functions subsequent to base pair formation with U6. Our data suggest that one such function is likely to be in tri-snRNP formation, when U5 joins the U4/U6 di-snRNP. We have identified two regions, the upper stem of the 3' stem-loop and the central domain, that promote tri-snRNP formation. In addition, the loop of the 3' stem-loop promotes di-snRNP formation, while the central domain and the 3'-terminal domain appear to antagonize di-snRNP formation.
Ansari,
SLU7 and a novel activity, SSF1, act during the PRP16-dependent step of yeast pre-mRNA splicing.
1995, Pubmed
Ansari,
SLU7 and a novel activity, SSF1, act during the PRP16-dependent step of yeast pre-mRNA splicing.
1995,
Pubmed
Aukema,
Small molecule inhibitors of yeast pre-mRNA splicing.
2009,
Pubmed
Ayadi,
Mutations within the yeast U4/U6 snRNP protein Prp4 affect a late stage of spliceosome assembly.
1997,
Pubmed
Berget,
U1, U2, and U4/U6 small nuclear ribonucleoproteins are required for in vitro splicing but not polyadenylation.
1986,
Pubmed
Bordonné,
Domains of yeast U4 spliceosomal RNA required for PRP4 protein binding, snRNP-snRNP interactions, and pre-mRNA splicing in vivo.
1990,
Pubmed
Brenner,
Assembly of Snu114 into U5 snRNP requires Prp8 and a functional GTPase domain.
2006,
Pubmed
Brow,
Spliceosomal RNA U6 is remarkably conserved from yeast to mammals.
1988,
Pubmed
Cheng,
Spliceosome assembly in yeast.
1987,
Pubmed
Fabrizio,
In vitro assembly of yeast U6 snRNP: a functional assay.
1989,
Pubmed
Ghetti,
Specificity of Prp24 binding to RNA: a role for Prp24 in the dynamic interaction of U4 and U6 snRNAs.
1995,
Pubmed
Guthrie,
Spliceosomal snRNAs.
1988,
Pubmed
He,
Mutations in U4atac snRNA, a component of the minor spliceosome, in the developmental disorder MOPD I.
2011,
Pubmed
Horowitz,
A U5 small nuclear ribonucleoprotein particle protein involved only in the second step of pre-mRNA splicing in Saccharomyces cerevisiae.
1993,
Pubmed
Hu,
Mutational analysis of Saccharomyces cerevisiae U4 small nuclear RNA identifies functionally important domains.
1995,
Pubmed
Hu,
The central region of the yeast U4 snRNA is not important for hammerhead catalysis, but may act as a domain spacer.
1995,
Pubmed
Lamond,
Spliceosome assembly involves the binding and release of U4 small nuclear ribonucleoprotein.
1988,
Pubmed
Leung,
Structure of the spliceosomal U4 snRNP core domain and its implication for snRNP biogenesis.
2011,
Pubmed
Mayas,
Exon ligation is proofread by the DExD/H-box ATPase Prp22p.
2006,
Pubmed
McGrail,
Mutation in the U2 snRNA influences exon interactions of U5 snRNA loop 1 during pre-mRNA splicing.
2006,
Pubmed
McGrail,
The U1, U2 and U5 snRNAs crosslink to the 5' exon during yeast pre-mRNA splicing.
2008,
Pubmed
McPheeters,
In vitro reconstitution of functional yeast U2 snRNPs.
1989,
Pubmed
Myslinski,
A phylogenetic study of U4 snRNA reveals the existence of an evolutionarily conserved secondary structure corresponding to 'free' U4 snRNA.
1991,
Pubmed
O'Keefe,
The invariant U5 snRNA loop 1 sequence is dispensable for the first catalytic step of pre-mRNA splicing in yeast.
1996,
Pubmed
Owczarzy,
IDT SciTools: a suite for analysis and design of nucleic acid oligomers.
2008,
Pubmed
Raghunathan,
A spliceosomal recycling factor that reanneals U4 and U6 small nuclear ribonucleoprotein particles.
1998,
Pubmed
Raghunathan,
RNA unwinding in U4/U6 snRNPs requires ATP hydrolysis and the DEIH-box splicing factor Brr2.
1998,
Pubmed
Shukla,
Domains of human U4atac snRNA required for U12-dependent splicing in vivo.
2002,
Pubmed
Ségault,
In vitro reconstitution of mammalian U2 and U5 snRNPs active in splicing: Sm proteins are functionally interchangeable and are essential for the formation of functional U2 and U5 snRNPs.
1995,
Pubmed
Vankan,
Domains of U4 and U6 snRNAs required for snRNP assembly and splicing complementation in Xenopus oocytes.
1990,
Pubmed
,
Xenbase
Vankan,
Roles of U4 and U6 snRNAs in the assembly of splicing complexes.
1992,
Pubmed
,
Xenbase
Vijayraghavan,
Mutations in conserved intron sequences affect multiple steps in the yeast splicing pathway, particularly assembly of the spliceosome.
1986,
Pubmed
Wersig,
Conserved domains of human U4 snRNA required for snRNP and spliceosome assembly.
1990,
Pubmed
Wersig,
Assembly and nuclear transport of the U4 and U4/U6 snRNPs.
1992,
Pubmed
,
Xenbase
Wersig,
Reconstitution of functional mammalian U4 small nuclear ribonucleoprotein: Sm protein binding is not essential for splicing in vitro.
1992,
Pubmed
Will,
In vitro reconstitution of mammalian U1 snRNPs active in splicing: the U1-C protein enhances the formation of early (E) spliceosomal complexes.
1996,
Pubmed
,
Xenbase
Wolff,
Reconstituted mammalian U4/U6 snRNP complements splicing: a mutational analysis.
1992,
Pubmed
Xu,
The PRP4 (RNA4) protein of Saccharomyces cerevisiae is associated with the 5' portion of the U4 small nuclear RNA.
1990,
Pubmed
Yean,
U4 small nuclear RNA dissociates from a yeast spliceosome and does not participate in the subsequent splicing reaction.
1991,
Pubmed