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Nucleic Acids Res
1996 Dec 01;2423:4614-23. doi: 10.1093/nar/24.23.4614.
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In vitro reconstitution of mammalian U1 snRNPs active in splicing: the U1-C protein enhances the formation of early (E) spliceosomal complexes.
Will CL, Rümpler S, Klein Gunnewiek J, van Venrooij WJ, Lührmann R.
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We have established an in vitro reconstitution/splicing complementation system which has allowed the investigation of the role of mammalian U1 snRNP components both in splicing and at the early stages of spliceosome formation. U1 snRNPs reconstituted from purified, native snRNP proteins and either authentic or in vitro transcribed U1 snRNA restored both early (E) splicing complex formation and splicing-activity to U1-depleted extracts. In vitro reconstituted U1 snRNPs possessing an m3G or ApppG cap were equally active in splicing, demonstrating that a physiological cap structure is not absolutely required for U1 function. However, the presence of an m7GpppG or GpppG cap was deleterious to splicing, most likely due to competition for the m7G cap binding proteins. No significant reduction in splicing or E complex formation was detected with U1 snRNPs reconstituted from U1 snRNA lacking the RNA binding sites of the U1-70K or U1-A protein (i.e., stem-loop I and II, respectively). Complementation studies with purified HeLa U1 snRNPs lacking subsets of the U1-specific proteins demonstrated a role for the U1-C, but not U1-A, protein in the formation and/or stabilization of early splicing complexes. Studies with recombinant U1-C protein mutants indicated that the N-terminal domain of U1-C is necessary and sufficient for the stimulation of E complex formation.
Bach,
Purification of small nuclear ribonucleoprotein particles with antibodies against modified nucleosides of small nuclear RNAs.
1990, Pubmed
Bach,
Purification of small nuclear ribonucleoprotein particles with antibodies against modified nucleosides of small nuclear RNAs.
1990,
Pubmed Branlant,
U2 RNA shares a structural domain with U1, U4, and U5 RNAs.
1982,
Pubmed Colot,
The yeast splicing factor Mud13p is a commitment complex component and corresponds to CBP20, the small subunit of the nuclear cap-binding complex.
1996,
Pubmed Dignam,
Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.
1983,
Pubmed Fischer,
An essential signaling role for the m3G cap in the transport of U1 snRNP to the nucleus.
1990,
Pubmed
,
Xenbase Frendewey,
Stepwise assembly of a pre-mRNA splicing complex requires U-snRNPs and specific intron sequences.
1985,
Pubmed Fu,
The superfamily of arginine/serine-rich splicing factors.
1995,
Pubmed Gunnewiek,
Homodimerization of the human U1 snRNP-specific protein C.
1995,
Pubmed Hamm,
Monomethylated cap structures facilitate RNA export from the nucleus.
1990,
Pubmed
,
Xenbase Hamm,
In vitro assembly of U1 snRNPs.
1987,
Pubmed
,
Xenbase Hamm,
Multiple domains of U1 snRNA, including U1 specific protein binding sites, are required for splicing.
1990,
Pubmed
,
Xenbase Heinrichs,
U1-specific protein C needed for efficient complex formation of U1 snRNP with a 5' splice site.
1990,
Pubmed Hilleren,
The amino-terminal domain of yeast U1-70K is necessary and sufficient for function.
1995,
Pubmed Izaurralde,
A nuclear cap binding protein complex involved in pre-mRNA splicing.
1994,
Pubmed
,
Xenbase Jamison,
U1 snRNP-ASF/SF2 interaction and 5' splice site recognition: characterization of required elements.
1995,
Pubmed Kohtz,
Protein-protein interactions and 5'-splice-site recognition in mammalian mRNA precursors.
1994,
Pubmed Lewis,
A nuclear cap-binding complex facilitates association of U1 snRNP with the cap-proximal 5' splice site.
1996,
Pubmed Mattaj,
Cap trimethylation of U snRNA is cytoplasmic and dependent on U snRNP protein binding.
1986,
Pubmed
,
Xenbase Melton,
Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter.
1984,
Pubmed Michaud,
A functional association between the 5' and 3' splice site is established in the earliest prespliceosome complex (E) in mammals.
1993,
Pubmed Michaud,
An ATP-independent complex commits pre-mRNA to the mammalian spliceosome assembly pathway.
1991,
Pubmed Mount,
The U1 small nuclear RNA-protein complex selectively binds a 5' splice site in vitro.
1983,
Pubmed Nelissen,
The association of the U1-specific 70K and C proteins with U1 snRNPs is mediated in part by common U snRNP proteins.
1994,
Pubmed Nelissen,
Zinc finger-like structure in U1-specific protein C is essential for specific binding to U1 snRNP.
1991,
Pubmed Patton,
Reconstitution of the U1 small nuclear ribonucleoprotein particle.
1987,
Pubmed Patton,
The Mr 70,000 protein of the U1 small nuclear ribonucleoprotein particle binds to the 5' stem-loop of U1 RNA and interacts with Sm domain proteins.
1988,
Pubmed Query,
A common RNA recognition motif identified within a defined U1 RNA binding domain of the 70K U1 snRNP protein.
1989,
Pubmed Rossi,
Involvement of U1 small nuclear ribonucleoproteins (snRNP) in 5' splice site-U1 snRNP interaction.
1996,
Pubmed Ruby,
An early hierarchic role of U1 small nuclear ribonucleoprotein in spliceosome assembly.
1988,
Pubmed Scherly,
Identification of the RNA binding segment of human U1 A protein and definition of its binding site on U1 snRNA.
1989,
Pubmed
,
Xenbase 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 Seraphin,
Identification of functional U1 snRNA-pre-mRNA complexes committed to spliceosome assembly and splicing.
1989,
Pubmed Siliciano,
5' splice site selection in yeast: genetic alterations in base-pairing with U1 reveal additional requirements.
1988,
Pubmed Sillekens,
cDNA cloning of the human U1 snRNA-associated A protein: extensive homology between U1 and U2 snRNP-specific proteins.
1987,
Pubmed Sillekens,
Human U1 snRNP-specific C protein: complete cDNA and protein sequence and identification of a multigene family in mammals.
1988,
Pubmed Sumpter,
In vitro reconstitution of U1 and U2 snRNPs from isolated proteins and snRNA.
1992,
Pubmed Wersig,
Reconstitution of functional mammalian U4 small nuclear ribonucleoprotein: Sm protein binding is not essential for splicing in vitro.
1992,
Pubmed Will,
Protein composition of mammalian spliceosomal snRNPs.
1993,
Pubmed Wolff,
Reconstituted mammalian U4/U6 snRNP complements splicing: a mutational analysis.
1992,
Pubmed Zhuang,
A compensatory base change in U1 snRNA suppresses a 5' splice site mutation.
1986,
Pubmed Zillmann,
Gel electrophoretic isolation of splicing complexes containing U1 small nuclear ribonucleoprotein particles.
1988,
Pubmed
