Structure of spliceosomal snRNPs and their role in pre-mRNA splicing
Biochimica et Biophysica Acta (BBA) - Gene Structure and ExpressionPublished 1 November 1990
Reinhard Lührmann, Berthold Kastner, M.A. Bach
Citations409
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Abstract
Synthese des Small nuclear Ribo Nucleoproteines (SnRNP) essentielles a l'epissage des premRNA nucleaires: composition, interactions proteines-proteines ou proteines-RNA, structures en microscopie electronique et role au niveau du spliceosome
Keywords
Biochemistry, Genetics and Molecular Biology
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A procedure for preparing extracts from nuclei of human tissue culture cells that directs accurate transcription initiation in vitro from class II promoters, including tRNA and Ad 2 VA, is developed.
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NatureAre snRNPs involved in splicing?
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Several lines of evidence are presented that suggest a direct involvement of snRNPs in the splicing of hnRNA molecules, including the observation that the nucleotide sequence at the 5′ end of U1 RNA exhibits extensive complementarity to those across splice junctions in hn RNA molecules.
Proceedings of the National Academy of SciencesAntibodies to small nuclear RNAs complexed with proteins are produced by patients with systemic lupus erythematosus
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It is argued that each of the six snRNAs exists in a separate small nuclear ribonucleoprotein (snRNP) complex with a total molecular weight of about 175,000.
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922 Citations1987Phillip A. Sharp
A general mechanism for the splicing of nuclear messenger RNA precursors in eukaryotic cells, which generates lariat RNAs possessing a branch site, seems related to the RNA-catalyzed reactions of self-splicing introns.
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761 Citations1986Yuan Zhuang, Alan M. Weiner
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CellNormal and mutant human β-globin pre-mRNAs are faithfully and efficiently spliced in vitro
674 Citations1984Adrian R. Krainer, Tom Maniatis +2 more
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The RNA binding domain of the 70K protein component of the U1 small nuclear ribonucleoprotein is defined and its aberrant electrophoretic migration is found to be due to a carboxy-terminal charged domain structurally similar to two Drosophila proteins that may regulate alternative pre-messenger RNA splicing.
NatureThe role of small nuclear ribonucleoprotein particles in pre-mRNA splicing
596 Citations1987Tom Maniatis, Robin Reed
A small set of distinctive short RNA molecules are found in the nuclei of all higher eukaryotic cells and yeast, in protein complexes known as 'small nuclear ribonucleoprotein particles', or snRNPs, which have been shown to be an integral part of the ' spliceosome'.
CellThe U1 small nuclear RNA-protein complex selectively binds a 5′ splice site in vitro
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CellU2 as well as U1 small nuclear ribonucleoproteins are involved in premessenger RNA splicing
573 Citations1985Douglas L. Black, Benoı̂t Chabot +1 more
Use of anti-(U2)RNP autoantibodies demonstrates that U2 snRNPs associate with the precursor RNA during in vitro splicing and possible interactions of the U2snRNP with the U1 snRNP and with the pre-mRNA during splicing are considered.
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555 Citations1980John Rogers, Randolph Wall
U-1 small nuclear RNA is proposed to be the recognition component of the nuclear RNA splicing enzyme and forms base pairs with both ends of an intron so as to align them for cutting and splicing.
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543 Citations1988Barbara Ruskin, Phillip D. Zamore +1 more
This work identifies a previously uncharacterized activity, U2AF, that is required for the U2 snRNP-branch point interaction and splicing complex formation and provides an explanation for the necessity of the 3' splice site region in U2SnRNP binding and, hence, the first step of splicing.
CellInteractions between small nuclear ribonucleoprotein particles in formation of spliceosomes
515 Citations1987Maria M. Konarska, Phillip A. Sharp
Electrophoretic separation of ribonucleoprotein particles in a nondenaturing gel was used to analyze the splicing of mRNA precursors and surprisingly, U4 snRNP was not part of the intron-containing complex, suggesting that U4/6 snR NP disassembles and assembles during splicing.
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511 Citations1987Roy D. Parker, Paul G. Siliciano +1 more
Recognition of the UACUAAC box relies, at least in part, on Watson-Crick base pairing with the yeast U2 analogue, and allele-specific suppression of biological and biochemical phenotypes was observed.
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451 Citations1990Susan Kass, Kazimierz Tyc +2 more
The identified U3 function is discussed with respect to the molecular basis of pre-rRNA recognition by the U3 snRNP, possible roles of U3 and other nucleolar snRNPs in rRNA processing, and the morphological organization of the nucleolus and the ribosomal transcription complex.
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CellMultiple factors including the small nuclear ribonucleoproteins U1 and U2 are necessary for Pre-mRNA splicing in vitro
443 Citations1985Adrian R. Krainer, Tom Maniatis
Six distinct factors necessary for pre-mRNA splicing in vitro are identified by selective inactivation and complementation studies, and by fractionation procedures.
NatureSpliceosomal RNA U6 is remarkably conserved from yeast to mammals
429 Citations1988David A. Brow, Christine Guthrie
An extensive base-pairing interaction with U4 snRNA is described; the destabilization of the U4/U6 complex seen during splicing thus requires a large conformational change.
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407 Citations1984Angela Krämer, Walter Keller +2 more
These results are in strong support of current models suggesting that a base-pairing interaction between the 5' terminus of the U1 snRNA and the5' splice site of a mRNA precursor is a prerequisite for proper splicing.
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402 Citations1988Gideon Dreyfuss, Maurice S. Swanson +1 more
Heterogeneous nuclear ribonucleoprotein particles, the structures that package hnRNA, are one of the major constituents of the nucleus and the discovery of a conserved RNA-binding domain and a RNP consensus sequence made possible.
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A new strategy to prepare snR NP-depleted splicing extracts using Saccharomyces cerevisiae cells conditionally expressing U1 or U2 snRNP is developed and it is demonstrated that these complexes are functional intermediates in spliceosome assembly and splicing.
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386 Citations1987Soo-Chen Cheng, John Abelson
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CellCap trimethylation of U snRNA is cytoplasmic and dependent on U snRNP protein binding
386 Citations1986Iain W. Mattaj
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381 Citations1989Phillip D. Zamore, M R Green
U2AF initiates mammalian spliceosome assembly by facilitating the ATP-dependent binding of U2 snRNP and binds rapidly at 4 degrees C in the absence of ATP.
Genes & DevelopmentIdentification and purification of a 62,000-dalton protein that binds specifically to the polypyrimidine tract of introns.
347 Citations1989Mariano A. García-Blanco, Sharon F. Jamison +1 more
Alterations in the polypyrimidine tract that reduce the binding of p62 yield a corresponding reduction in the efficiency of formation of a U2 snRNP/pre-mRNA complex and splicing.
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334 Citations1983Richard A. Padgett, Stephen M. Mount +2 more
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313 Citations1987Charles C. Query
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ScienceThe 3′ Splice Site of Pre-Messenger RNA Is Recognized by a Small Nuclear Ribonucleoprotein
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306 Citations1989Robin Reed
A model pre-mRNA substrate was used to carry out a detailed investigation of the functional organization of sequences at the 3' end of mammalian introns and revealed a difference in the sequence requirements for the first and second steps of the splicing reaction.
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288 Citations1989Yuan Zhuang, Alan M. Weiner
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274 Citations1986Jamal Tazi, Christine Alibert +5 more
Although the technique used here shows that the binding specificity is an intrinsic property of IBP and does not depend on snRNA-pre-mRNA interactions, it comigrates with U5 snRNP and is immunoprecipitated by anti-Sm antibody, which strongly suggests that IBP belongs to U5SnRNP.
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268 Citations1989Christopher W. J. Smith, Elena B. Porro +2 more
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The results suggest that interaction of the two splice site regions occurs at an early stage of spliceosome formation and is probably mediated by U1 snRNP and perhaps other factors.
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239 Citations1985Barbara Ruskin, J. M. Greene +1 more
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The EMBO JournalCloning of the human cDNA for the U1 RNA‐associated 70K protein.
235 Citations1986H. Theissen, Michael Etzerodt +6 more
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The RNA-RNA interaction may be sufficient to maintain U4 and U6 RNAs in the same snRNP particle, consistent with the existence of specific base-pairing between the two RNAs.
CellPre-mRNA splicing in vitro requires intact U4/U6 small nuclear ribonucleoprotein
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All four abundant snRNPs, containing U1, U2, U5, or U4 and U6 RNAs, are now implicated in pre-mRNA splicing.
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The EMBO JournalcDNA cloning of the human U1 snRNA‐associated A protein: extensive homology between U1 and U2 snRNP‐specific proteins.
202 Citations1987Peter Sillekens, W J Habets +2 more
Sequence comparison with the recently cloned U2 snRNP‐specific B′ protein revealed two extremely homologous regions located in the carboxy‐terminal and amino‐ terminal part of the proteins, which indicates that proteins A and B′, although located in different snR NP particles, may have identical functions.
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201 Citations1988Ram Reddy, Harris Busch
All eukaryotic cells contain multiple small nuclear RNAs, designated U- SnRNAs, which account for about 1% total mammalian cellular RNA; of these, the U1 to U8 snRNAs have been sively characterized.
CellU1, U2, and U4/U6 small nuclear ribonucleoproteins are required for in vitro splicing but not polyadenylation
192 Citations1986Susan M. Berget, Barbara Robberson
The involvement of multiple snRNPs in the splicing of simple precursor RNAs suggests that the spliceosome is a large complex assembly consisting of multiplesnRNPs whose activity is dependent on the structural integrity of the individual U RNAs.
Proceedings of the National Academy of SciencesSpliceosome assembly involves the binding and release of U4 small nuclear ribonucleoprotein.
189 Citations1988Angus I. Lamond, Maria M. Konarska +2 more
A pathway of spliceosome assembly was deduced that has at least three stages and U2 small nuclear ribonucleoprotein alone binds to sequences of mRNA upstream of the 3' splice site.
Molecular and Cellular BiologyGel electrophoretic isolation of splicing complexes containing U1 small nuclear ribonucleoprotein particles.
189 Citations1988Martin Zillmann, Maria L. Zapp +1 more
Proceedings of the National Academy of SciencesGamma-monomethyl phosphate: a cap structure in spliceosomal U6 small nuclear RNA.
187 Citations1989Ravi Singh, R Reddy
The cap structure of human U6 snRNA is characterized and shows that the gamma phosphate of the 5' guanosine triphosphate is methylated, distinct from all other cap structures characterized thus far.
Trends in GeneticsOn/off regulation of gene expression at the level of splicing
184 Citations1988Paul M. Bingham, Tze-Bin Chou +2 more
The past two years have seen the discovery of three independent cases in which expression of a eukaryotic protein gene product is turned on and off by controlling splicing events necessary to produce the corresponding mRNA.
Journal of Biological ChemistryThe capped U6 small nuclear RNA is transcribed by RNA polymerase III.
184 Citations1987R Reddy, D Henning +3 more
U6 RNA, though capped on its 5'-end, is transcribed by RNA polymerase III; this is the first report of a capped RNA molecule synthesized by RNA Polymerase III.
CellIdentification of a yeast snRNP protein and detection of snRNP-snRNP interactions
183 Citations1987Marie Lossky, Gordon J. Anderson +2 more
Immunological procedures have been used to demonstrate that the RNA8 protein is in stable association with the small nuclear RNAs snR7L andSnR7S, which are also known to be required for splicing and which are present in spliceosomal complexes.
CellA putative ATP binding protein influences the fidelity of branchpoint recognition in yeast splicing
182 Citations1990Sean M. Burgess, Joseph Couto +1 more
It is proposed that PRP16 is an excellent candidate for mediating one of the many ATP-requiring steps of spliceosome assembly and that accuracy of branchpoint recognition may be coupled to ATP binding and/or hydrolysis.
Proceedings of the National Academy of SciencesUACUAAC is the preferred branch site for mammalian mRNA splicing.
181 Citations1989Yuan Zhuang, Alisa M. Goldstein +1 more
The results suggest that in mammals U2 snRNA can form base pairs with the branch site and the interaction between U2 and the branchsite can be augmented or replaced by an interaction between the spliceosome and some other element of the intron or exons, perhaps the conserved polypyrimidine tract located immediately upstream from the 3' splice site.
Journal of Molecular BiologyStructural organization of ribonucleoproteins containing small nuclear RNAs from HeLa cells
180 Citations1982Jean‐Pierre Liautard, Joannés Sri-Widada +2 more
CellSpecific and stable intron-factor interactions are established early during in vitro pre-mRNA splicing
180 Citations1985Barbara Ruskin, Michael R. Green
Results suggest that selection of the 3' splice site accompanied by the association of a factor with the branch point may be the initial step in mammalian pre-mRNA splicing.
Genes & DevelopmentA mutational analysis of spliceosome assembly: evidence for splice site collaboration during spliceosome formation.
174 Citations1987Angus I. Lamond, Maria M. Konarska +1 more
Surprisingly, the analysis of double mutants, with sequence alterations at both the 5' and 3' splice sites, revealed that the 5- splice site genotype was important for the efficient formation of a U2 snRNA-containing alpha complex at the 3'splice site.
The EMBO JournalPurification of the individual snRNPs U1, U2, U5 and U4/U6 from HeLa cells and characterization of their protein constituents.
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CellAntisense probing of the human U4U6 snRNP with biotinylated 2′-OMe RNA oligonucleotides
169 Citations1989Benjamin J. Blencowe, Brian S. Sproat +3 more
The results suggest that U4 snRNP is not released during spliceosome assembly, and that Pre-mRNA and splicing products could also be affinity-selected through binding of the oligonucleotides to U4/U6 snRNPs in splicing complexes.
ScienceTwo Domains of Yeast U6 Small Nuclear RNA Required for Both Steps of Nuclear Precursor Messenger RNA Splicing
169 Citations1990Patrizia Fabrizio, John Abelson
In both domains, the positions of base changes that block the second step of splicing correspond exactly to the site of insertion of pre-mRNA-type introns into the U6 gene of two yeast species, providing a possible explanation for the mechanism of how these introns originated and adding further evidence for the proposed catalytic role of U6 RNA.
CellAn essential yeast snRNA with a U5-like domain is required for splicing in vivo
169 Citations1987Bruce K. Patterson, Christine Guthrie
It is shown that mRNA splicing in yeast requires the function of a U5-like snRNA, and a 70 nucleotide domain within snR7 is identified with limited sequence-specific but striking structural homology to the mammalian snRNA U5.
Proceedings of the National Academy of SciencesSplicing of adenovirus RNA in a cell-free transcription system.
160 Citations1983Richard A. Padgett, Stephen Hardy +1 more
A soluble whole-cell extract prepared accurately from HeLa cells splices 2-3% of the RNA transcribed from a DNA template containing the first and second leader exons of late adenovirus RNA.
Nucleic Acids ResearchStructure and organization of the gene coding for the DNA binding protein of adenovirus type 5
154 Citations1981W. Kruijer, F.M.A. van Schaik +1 more
From the nucleotide sequence of the DBP gene it can be derived that DBP contains 529 amino acid residues and has an actual molecular weight of 59,049 daltons.
Proceedings of the National Academy of Sciences20S small nuclear ribonucleoprotein U5 shows a surprisingly complex protein composition.
147 Citations1989M.A. Bach, G. Winkelmann +1 more
Nucleic Acids ResearchThe human U1-70K snRNP protein: cDNA cloning, chromosomal localization, expression, alternative splicing and RNA-binding
146 Citations1987Richard A. Spritz, Kathleen M. Strunk +4 more
Comparison of the cDNA sequences indicates that there are multiple subclasses of mRNA that arise by alternative pre-mRNA splicing of at least four alternative exon segments, which suggests that multiple forms of the hU1-70K protein may exist, possibly with different functions in vivo.
Molecular and General Genetics MGGDetermination of the location of proteins L14, L17, L18, L19, L22 and L23 on the surface of the 50S ribosomal subunit of Escherichia coli by immune electron microscopy
145 Citations1974Gilbert Tischendorf, Heinz Zeichhardt +1 more
The results allowed the proposal of a three-dimensional model of the 50S subunit with the location of six different ribosomal proteins as illustrated in Fig. 13.
The EMBO JournalPurification of snRNPs U1, U2, U4, U5 and U6 with 2,2,7‐trimethylguanosine‐specific antibody and definition of their constituent proteins reacting with anti‐Sm and anti‐(U1)RNP antisera.
143 Citations1983Peter Bringmann, Jutta Rinke +3 more
Six of the snRNP polypeptides, characterized by apparent mol.
Molecular and Cellular BiologyMultiple interactions between the splicing substrate and small nuclear ribonucleoproteins in spliceosomes.
143 Citations1987Benoı̂t Chabot, Joan A. Steitz
Proceedings of the National Academy of SciencesAdditional low-abundance human small nuclear ribonucleoproteins: U11, U12, etc.
141 Citations1988K. A. Montzka, J A Steitz
Two-dimensional gel fractionation has revealed the existence of a number of additional species of HeLa cell small RNAs that have 5' trimethylguanosine cap structures and are bound by proteins containing Sm epitopes, belonging to the Sm class of small nuclear ribonucleoproteins (snRNPs), whose best-known members are the four highly abundant particles required for pre-mRNA splicing.
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141 Citations1988Iain W. Mattaj
The purpose of this summary is to present extensively the conclusions of this chapter, and to point out some areas where further research is required.
ScienceU1-Specific Protein C needed for Efficient Complex Formation of U1 snRNP with a 5′ Splice Site
137 Citations1990Volker Heinrichs, M.A. Bach +2 more
The data suggest that protein C may potentiate the base-pair interaction between U1 RNA and the 5' splice site, which is important for the formation of the snRNP-pre-mRNA complex.
Proceedings of the National Academy of SciencesMolecular cloning of a cDNA encoding the human Sm-D autoantigen.
135 Citations1988Luis A. Rokeach, Jeanne A. Haselby +1 more
A cDNA clone isolated from a human B-lymphocyte cDNA library contains an open reading frame (ORF) capable of encoding a 13,282-dalton polypeptide and reveals a (Gly-Arg)9 repeated motif at the C terminus which may constitute one of the Sm-D immunoreactive determinants.
Molecular and Cellular BiologyPRP4: a protein of the yeast U4/U6 small nuclear ribonucleoprotein particle.
133 Citations1989Josette Banroques, John Abelson
Genes & DevelopmentIn vitro assembly of yeast U6 snRNP: a functional assay.
133 Citations1989Patrizia Fabrizio, David S. McPheeters +1 more
An in vitro system for reconstituting the functional U6 small ribonucleoprotein (snRNP) and shows that large deletions produce molecules that are unable to restore splicing activity in vitro and cannot interact with the endogenous U4 snRNA or form a mature spliceosome.
Journal of Molecular BiologyLocalization of a base-paired interaction between small nuclear RNAs U4 and U6 in intact U4/U6 ribonucleoprotein particles by psoralen cross-linking
132 Citations1985Jutta Rinke, Bernd Appel +2 more
The results indicate that proteins are not required to stabilize the mutual interactions between both RNAs, once they exist, and suggest, however, that proteins may well be needed for exposing the complementary RNA regions for proper intermolecular base-pairing in the course of the assembly of the U4/U6 RNP complex from isolated RNAs.
The EMBO JournalDomains of U4 and U6 snRNAs required for snRNP assembly and splicing complementation in Xenopus oocytes.
129 Citations1990P. Vankan, C. McGuigan +1 more
Combinations of mutants in the U4 and U6 interaction domain are used to show that there are not only requirements for base complementarity but also for specific sequences in these regions.
Proceedings of the National Academy of SciencesPRP5: a helicase-like protein required for mRNA splicing in yeast.
127 Citations1990Gloria Dalbadie‐McFarland, John Abelson
The possible roles for an RNA helicase in splicing and for the activity of the PRP5 protein are discussed.
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127 Citations1988Gideon Dreyfuss, L. Philipson +1 more
Recent work from numerous laboratories has led to the discovery of many additional RNP particles that are involved in an unexpected variety of cellular processes including protein targeting, priming of mitochondrial DNA synthesis, and transcription termination.
Molecular and Cellular BiologyU5 small nuclear ribonucleoprotein: RNA structure analysis and ATP-dependent interaction with U4/U6.
124 Citations1989Douglas L. Black, A L Pinto
CellSmall nuclear ribonucleoprotein particle assembly in vivo: Demonstration of a 6S RNA-free core precursor and posttranslational modification
123 Citations1985David E. Fisher, Gregory E. Conner +3 more
Results indicate that assembly of the U RNA-containing snRNPs is a multistep process involving prior assembly of an RNA-free 6S core particle and a posttranslational modification of one of theU1 snRNP-specific peptides has been observed, and the kinetics of this process indicates that the modification occurs after particle assembly.
CellPurification and visualization of native spliceosomes
122 Citations1988Robin Reed, Jack D. Griffith +1 more
Evidence that these particles are spliceosomes is their sensitivity to micrococcal nuclease, their ATP-dependent assembly, and their immunoprecipitation with a trimethyl cap monoclonal antibody.
Nucleic Acids ResearchXenopus tropicalis U6 snRNA genes transcribed by Pot III contain the upstream promoter elements used by pol II dependent U snRNA genes
116 Citations1987Alain Krol, Philippe Carbon +2 more
It is proposed that U6 RNAs are synthesized by a specialized transcription complex consisting of RNA polymerase III and transcription factors, some of which are very likely shared withRNA polymerase II promoters.
CellProbing the structure and function of U2 snRNP with antisense oligonucleotides made of 2′-OMe RNA
115 Citations1989Angus I. Lamond, Brian S. Sproat +2 more
It is shown that antisense 2'-OMe RNA oligonucleotides bind efficiently and specifically to U2 snRNP and demonstrated that masking of two separate regions of U2snRNA can inhibit splicing by affecting different steps in the spliceosome assembly pathway.
ScienceDifferential Control of U1 Small Nuclear RNA Expression During Mouse Development
115 Citations1985Elsebet Lund, Brenda Kahan +1 more
The differential accumulation of embryonic U1 RNA's may influence the pattern of gene expression during early development and differentiation.
Genes & DevelopmentPresplicing complex formation requires two proteins and U2 snRNP.
113 Citations1988A. Krämer
Six fractions derived from a HeLa cell nuclear extract are necessary for the generation of spliced mRNA in vitro and their role in the formation of splicing complexes was analyzed by electrophoresis in native polyacrylamide gels.
Nucleic Acids ResearchPre-mRNA splicing by complementation with purified human UI, U2, U4/U6 and U5 snRNPs
112 Citations1988Adrian R. Krainer
Biochemical complementation studies show that the purified snRNPs are active in pre-mRNA splicing, but only in the presence of additional non-snRNP protein factors.
Trends in Biochemical Sciences‘Snurpogenesis’: The transcription and assembly of U snRNP components
103 Citations1989Huw D. Parry, Daniel Scherly +1 more
Assembly of the snRNAs with proteins to form snRNP particles has been studied in vivo and in vitro, and new in vitro systems promise to advance the understanding of the assembly process still further.
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