Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Site Tools

  • AAAS
  • Subscribe
  • Feedback

Site Search

Search Advanced

Science 24 December 1993:
Vol. 262. no. 5142, pp. 1982 - 1988
DOI: 10.1126/science.8266093
Prev | Table of Contents | Next

Articles

Science, Vol 262, Issue 5142, 1982-1988
Copyright © 1993 by American Association for the Advancement of Science

articles

Mutations in U6 snRNA that alter splice site specificity: implications for the active site

CF Lesser and C Guthrie

Medical Scientist Training Program, University of California, San Francisco 94143.

What determines the precise sites of cleavage in the two transesterification reactions of messenger RNA (mRNA) splicing is a major unsolved question. Mutation of the invariant G (guanosine) at position 5 of 5' splice sites in Saccharomyces cerevisiae introns activates cleavage at nearby aberrant sites. A genetic approach was used to test the hypothesis that a base-pairing interaction between the 5' splice site and the invariant ACAGAG sequence of U6 is a determinant of 5' splice site choice. Mutations in U6 or the intron (or both) that were predicted to stabilize the interaction suppressed aberrant cleavage and increased normal cleavage. In addition, a mutation in the ACAGAG sequence suppressed mutations of the 3' splice site dinucleotide. These data can fit a model for the spliceosomal active site comprised of a set of RNA-RNA interactions between the intron, U2 and U6.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
A DNA damage signal activates and derepresses exon inclusion in Drosophila TAF1 alternative splicing.
M. S. Marengo and D. A. Wassarman (2008)
RNA 14, 1681-1695
   Abstract »    Full Text »    PDF »
The U1, U2 and U5 snRNAs crosslink to the 5' exon during yeast pre-mRNA splicing.
J. C. McGrail and R. T. O'Keefe (2008)
Nucleic Acids Res. 36, 814-825
   Abstract »    Full Text »    PDF »
Three essential and conserved regions of the group II intron are proximal to the 5'-splice site.
A. de Lencastre and A. M. Pyle (2008)
RNA 14, 11-24
   Abstract »    Full Text »    PDF »
Protein-free spliceosomal snRNAs catalyze a reaction that resembles the first step of splicing.
S. Valadkhan, A. Mohammadi, C. Wachtel, and J. L. Manley (2007)
RNA 13, 2300-2311
   Abstract »    Full Text »    PDF »
Structural RNAs of known and unknown function identified in malaria parasites by comparative genomics and RNA analysis.
K. Chakrabarti, M. Pearson, L. Grate, T. Sterne-Weiler, J. Deans, J. P. Donohue, and M. Ares Jr (2007)
RNA 13, 1923-1939
   Abstract »    Full Text »    PDF »
In vitro and in silico analysis reveals an efficient algorithm to predict the splicing consequences of mutations at the 5' splice sites.
K. Sahashi, A. Masuda, T. Matsuura, J. Shinmi, Z. Zhang, Y. Takeshima, M. Matsuo, G. Sobue, and K. Ohno (2007)
Nucleic Acids Res. 35, 5995-6003
   Abstract »    Full Text »    PDF »
Aberrant 5' splice sites in human disease genes: mutation pattern, nucleotide structure and comparison of computational tools that predict their utilization.
E. Buratti, M. Chivers, J. Kralovicova, M. Romano, M. Baralle, A. R. Krainer, and I. Vorechovsky (2007)
Nucleic Acids Res. 35, 4250-4263
   Abstract »    Full Text »    PDF »
A Sequence Motif in the Simian Virus 40 (SV40) Early Core Promoter Affects Alternative Splicing of Transcribed mRNA.
E. Gendra, D. F. Colgan, B. Meany, and M. M. Konarska (2007)
J. Biol. Chem. 282, 11648-11657
   Abstract »    Full Text »    PDF »
Dissection of Prp8 protein defines multiple interactions with crucial RNA sequences in the catalytic core of the spliceosome.
I. A. TURNER, C. M. NORMAN, M. J. CHURCHER, and A. J. NEWMAN (2006)
RNA 12, 375-386
   Abstract »    Full Text »    PDF »
The Prp19-associated Complex Is Required for Specifying Interactions of U5 and U6 with Pre-mRNA during Spliceosome Activation.
S.-P. Chan and S.-C. Cheng (2005)
J. Biol. Chem. 280, 31190-31199
   Abstract »    Full Text »    PDF »
The Isy1p component of the NineTeen Complex interacts with the ATPase Prp16p to regulate the fidelity of pre-mRNA splicing.
T. Villa and C. Guthrie (2005)
Genes & Dev. 19, 1894-1904
   Abstract »    Full Text »    PDF »
Determinants of the inherent strength of human 5' splice sites.
X. ROCA, R. SACHIDANANDAM, and A. R. KRAINER (2005)
RNA 11, 683-698
   Abstract »    Full Text »    PDF »
{psi}35 in the Branch Site Recognition Region of U2 Small Nuclear RNA Is Important for Pre-mRNA Splicing in Saccharomyces cerevisiae.
C. Yang, D. S. McPheeters, and Y.-T. Yu (2005)
J. Biol. Chem. 280, 6655-6662
   Abstract »    Full Text »    PDF »
Multiple functions for the invariant AGC triad of U6 snRNA.
A. K. HILLIKER and J. P. STALEY (2004)
RNA 10, 921-928
   Abstract »    Full Text »    PDF »
Ordered Partitioning Reveals Extended Splice-Site Consensus Information.
M. Weir and M. Rice (2004)
Genome Res. 14, 67-78
   Abstract »    Full Text »    PDF »
The Prp19p-Associated Complex in Spliceosome Activation.
S.-P. Chan, D.-I Kao, W.-Y. Tsai, and S.-C. Cheng (2003)
Science 302, 279-282
   Abstract »    Full Text »    PDF »
Mutagenesis Suggests Several Roles of Snu114p in Pre-mRNA Splicing.
C. Bartels, H. Urlaub, R. Luhrmann, and P. Fabrizio (2003)
J. Biol. Chem. 278, 28324-28334
   Abstract »    Full Text »    PDF »
Spatial Organization of Protein-RNA Interactions in the Branch Site-3' Splice Site Region during pre-mRNA Splicing in Yeast.
D. S. McPheeters and P. Muhlenkamp (2003)
Mol. Cell. Biol. 23, 4174-4186
   Abstract »    Full Text »    PDF »
The U1 snRNP Base Pairs with the 5' Splice Site within a Penta-snRNP Complex.
H. Malca, N. Shomron, and G. Ast (2003)
Mol. Cell. Biol. 23, 3442-3455
   Abstract »    Full Text »    PDF »
Characterization of U4 and U6 interactions with the 5' splice site using a S. cerevisiae in vitro trans-splicing system.
T. L. Johnson and J. Abelson (2001)
Genes & Dev. 15, 1957-1970
   Abstract »    Full Text »    PDF »
A Novel Yeast U2 snRNP Protein, Snu17p, Is Required for the First Catalytic Step of Splicing and for Progression of Spliceosome Assembly.
A. Gottschalk, C. Bartels, G. Neubauer, R. Lührmann, and P. Fabrizio (2001)
Mol. Cell. Biol. 21, 3037-3046
   Abstract »    Full Text »
Suppressors of a Cold-Sensitive Mutation in Yeast U4 RNA Define Five Domains in the Splicing Factor Prp8 That Influence Spliceosome Activation.
A. N. Kuhn and D. A. Brow (2000)
Genetics 155, 1667-1682
   Abstract »    Full Text »
Interaction of the yeast DExH-box RNA helicase Prp22p with the 3' splice site during the second step of nuclear pre-mRNA splicing.
D. S. McPheeters, B. Schwer, and P. Muhlenkamp (2000)
Nucleic Acids Res. 28, 1313-1321
   Abstract »    Full Text »    PDF »
The Allele-Specific Suppressor sup-39 Alters Use of Cryptic Splice Sites in Caenorhabditis elegans.
A. B. Roller, D. C. Hoffman, and A. M. Zahler (2000)
Genetics 154, 1169-1179
   Abstract »    Full Text »
Role of adenine functional groups in the recognition of the 3'-splice-site AG during the second step of pre-mRNA splicing.
R. K. Gaur, L. Beigelman, P. Haeberli, and T. Maniatis (2000)
PNAS 97, 115-120
   Abstract »    Full Text »    PDF »
Extensive Genetic Interactions Between PRP8 and PRP17/CDC40, Two Yeast Genes Involved in Pre-mRNA Splicing and Cell Cycle Progression.
S. Ben-Yehuda, C. S. Russell, I. Dix, J. D. Beggs, and M. Kupiec (2000)
Genetics 154, 61-71
   Abstract »    Full Text »
Allele-specific genetic interactions between Prp8 and RNA active site residues suggest a function for Prp8 at the catalytic core of the spliceosome.
C. A. Collins and C. Guthrie (1999)
Genes & Dev. 13, 1970-1982
   Abstract »    Full Text »
Functional interactions of Prp8 with both splice sites at the spliceosomal catalytic center.
M. Siatecka, J. L. Reyes, and M. M. Konarska (1999)
Genes & Dev. 13, 1983-1993
   Abstract »    Full Text »
Pseudouridine Mapping in the Saccharomyces cerevisiae Spliceosomal U Small Nuclear RNAs (snRNAs) Reveals that Pseudouridine Synthase Pus1p Exhibits a Dual Substrate Specificity for U2 snRNA and tRNA.
S. Massenet, Y. Motorin, D. L. J. Lafontaine, E. C. Hurt, H. Grosjean, and C. Branlant (1999)
Mol. Cell. Biol. 19, 2142-2154
   Abstract »    Full Text »    PDF »
Detection of a Novel ATP-Dependent Cross-Linked Protein at the 5' Splice Site-U1 Small Nuclear RNA Duplex by Methylene Blue-Mediated Photo-Cross-Linking.
Z.-R. Liu, B. Sargueil, and C. W. J. Smith (1998)
Mol. Cell. Biol. 18, 6910-6920
   Abstract »    Full Text »
The Pre-mRNA 5' Cap Determines Whether U6 Small Nuclear RNA Succeeds U1 Small Nuclear Ribonucleoprotein Particle at 5' Splice Sites.
L. O'Mullane and I. C. Eperon (1998)
Mol. Cell. Biol. 18, 7510-7520
   Abstract »    Full Text »
The Human U5-220kD Protein (hPrp8) Forms a Stable RNA-Free Complex with Several U5-Specific Proteins, Including an RNA Unwindase, a Homologue of Ribosomal Elongation Factor EF-2, and a Novel WD-40 Protein.
T. Achsel, K. Ahrens, H. Brahms, S. Teigelkamp, and R. Lührmann (1998)
Mol. Cell. Biol. 18, 6756-6766
   Abstract »    Full Text »
Temperature-Sensitive Splicing in the Floral Homeotic Mutant apetala3-1.
R. W. M. Sablowski and E. M. Meyerowitz (1998)
PLANT CELL 10, 1453-1464
   Abstract »    Full Text »
Processing of the Intron-Encoded U18 Small Nucleolar RNA in the Yeast Saccharomyces cerevisiae Relies on Both Exo- and Endonucleolytic Activities.
T. Villa, F. Ceradini, C. Presutti, and I. Bozzoni (1998)
Mol. Cell. Biol. 18, 3376-3383
   Abstract »    Full Text »
Synthetic Lethality of Yeast slt Mutations with U2 Small Nuclear RNA Mutations Suggests Functional Interactions between U2 and U5 snRNPs That Are Important for Both Steps of Pre-mRNA Splicing.
D. Xu, D. J. Field, S.-J. Tang, A. Moris, B. P. Bobechko, and J. D. Friesen (1998)
Mol. Cell. Biol. 18, 2055-2066
   Abstract »    Full Text »
Site-specific crosslinking of mammalian U11 and U6atac to the 5' splice site of an AT-AC intron.
Y.-T. Yu and J. A. Steitz (1997)
PNAS 94, 6030-6035
   Abstract »    Full Text »    PDF »
Sequence Divergence Associated with Species-specific Splicing of the Nonmuscle beta-Tropomyosin Alternative Exon.
A.-M. Pret and M. Y. Fiszman (1996)
J. Biol. Chem. 271, 11511-11517
   Abstract »    Full Text »    PDF »
U1 snRNA promotes the selection of nearby 5' splice sites by U6 snRNA in mammalian cells..
D Y Hwang and J B Cohen (1996)
Genes & Dev. 10, 338-350
   Abstract »    PDF »
A U6 snRNA:pre-mRNA interaction can be rate-limiting for U1-independent splicing..
J D Crispino and P A Sharp (1995)
Genes & Dev. 9, 2314-2323
   Abstract »    PDF »
In Vitro Splicing Deficiency Induced by a C to T Mutation at Position -3 in the Intron 10 Acceptor Site of the Phenylalanine Hydroxylase Gene in a Patient with Phenylketonuria.
J. Jaruzelska, V. Abadie, Y. d'Aubenton-Carafa, E. Brody, A. Munnich, and J.ël. Marie (1995)
J. Biol. Chem. 270, 20370-20375
   Abstract »    Full Text »    PDF »
Disruption of Base-paired U4[IMAGE]U6 Small Nuclear RNAs Induced by Mammalian Heterogeneous Nuclear Ribonucleoprotein C Protein.
T. Forné, F. Rossi, E. Labourier, E. Antoine, G. Cathala, C. Brunel, and J. Tazi (1995)
J. Biol. Chem. 270, 16476-16481
   Abstract »    Full Text »    PDF »
A novel U2-U6 snRNA structure is necessary for mammalian mRNA splicing..
J S Sun and J L Manley (1995)
Genes & Dev. 9, 843-854
   Abstract »    PDF »
SR proteins can compensate for the loss of U1 snRNP functions in vitro..
W Y Tarn and J A Steitz (1994)
Genes & Dev. 8, 2704-2717
   Abstract »    PDF »
Complementation by SR proteins of pre-mRNA splicing reactions depleted of U1 snRNP.
J. Crispino, B. Blencowe, and P. Sharp (1994)
Science 265, 1866-1869
   Abstract »    PDF »
U4/U5/U6 snRNP recognizes the 5' splice site in the absence of U2 snRNP..
B B Konforti and M M Konarska (1994)
Genes & Dev. 8, 1962-1973
   Abstract »    PDF »
The prespliceosome components SAP 49 and SAP 145 interact in a complex implicated in tethering U2 snRNP to the branch site..
P Champion-Arnaud and R Reed (1994)
Genes & Dev. 8, 1974-1983
   Abstract »    PDF »
The influenza virus NS1 protein: a novel inhibitor of pre-mRNA splicing..
Y Lu, X Y Qian, and R M Krug (1994)
Genes & Dev. 8, 1817-1828
   Abstract »    PDF »
A group III intron is formed from domains of two individual group II introns..
L Hong and R B Hallick (1994)
Genes & Dev. 8, 1589-1599
   Abstract »    PDF »
A tertiary interaction in the Tetrahymena intron contributes to selection of the 5' splice site..
W D Downs and T R Cech (1994)
Genes & Dev. 8, 1198-1211
   Abstract »    PDF »
Randomization-selection analysis of snRNAs in vivo: evidence for a tertiary interaction in the spliceosome..
H D Madhani and C Guthrie (1994)
Genes & Dev. 8, 1071-1086
   Abstract »    PDF »
Guides to the heart of the spliceosome.
J. Wise (1993)
Science 262, 1978-1979
   PDF »
The U5 and U6 small nuclear RNAs as active site components of the spliceosome.
E. Sontheimer and J. Steitz (1993)
Science 262, 1989-1996
   Abstract »    PDF »
Involvement of U6 snRNA in 5' splice site selection.
S Kandels-Lewis and B Seraphin (1993)
Science 262, 2035-2039
   Abstract »    PDF »
Functional Analyses of Positions across the 5' Splice Site of the Trypanosomatid Spliced Leader RNA. IMPLICATIONS FOR BASE-PAIR INTERACTION WITH U5 AND U6 snRNAs.
Y.-x. Xu, L. Liu, and S. Michaeli (2000)
J. Biol. Chem. 275, 27883-27892
   Abstract »    Full Text »    PDF »


ADVERTISEMENT
Click Me!

ADVERTISEMENT
Click Me!

To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)