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Science 25 September 1992:
Vol. 257. no. 5078, pp. 1918 - 1925
DOI: 10.1126/science.1411506
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Articles

Science, Vol 257, Issue 5078, 1918-1925
Copyright © 1992 by American Association for the Advancement of Science

articles

Interactions of small nuclear RNA's with precursor messenger RNA during in vitro splicing

DA Wassarman and JA Steitz

Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, Haven, CT 06536.

Precursor messenger RNA splicing requires multiple factors including U1, U2, U4, U5, and U6 small nuclear RNA's. The crosslinking reagent psoralen was used to analyze the interactions of these RNA's with an adenovirus precursor messenger RNA in HeLa nuclear extract. An endogenous U2-U4-U6 crosslinkable complex dissociated upon incubation with precursor messenger RNA. During splicing, U1, U2, U5, and U6 became crosslinked to precursor messenger RNA and U2, U5, and U6 became crosslinked to excised lariat intron. U2 also formed a doubly crosslinked complex with U6 and precursor messenger RNA. The U1, U5, and U6 crosslinks to the precursor messenger RNA mapped to intron sequences near the 5' splice site, whereas the U2 crosslink mapped to the branch site. The kinetics of crosslink formation and disappearance delineates a temporal pathway for the action of small RNA's in the spliceosome. Potential base pairing interactions between conserved sequences in the small nuclear RNA's and precursor messenger RNA at the sites of crosslinking suggest that the 5' splice site is defined in several steps prior to the first cleavage event.


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U4/U5/U6 snRNP recognizes the 5' splice site in the absence of U2 snRNP..
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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
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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
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Upstream introns influence the efficiency of final intron removal and RNA 3'-end formation..
D Nesic and L E Maquat (1994)
Genes & Dev. 8, 363-375
   Abstract »    PDF »
A stem/loop in U6 RNA defines a conformational switch required for pre-mRNA splicing..
D M Fortner, R G Troy, and D A Brow (1994)
Genes & Dev. 8, 221-233
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Catalytic activity of an RNA domain derived from the U6-U4 RNA complex.
J. Yang, R Cedergren, and B Nadal-Ginard (1994)
Science 263, 77-81
   Abstract »    PDF »
Alternative pre-mRNA splicing: factors involved in splice site selection.
P. Norton (1994)
J. Cell Sci. 107, 1-7
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Guides to the heart of the spliceosome.
J. Wise (1993)
Science 262, 1978-1979
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Mutations in U6 snRNA that alter splice site specificity: implications for the active site.
C. Lesser and C Guthrie (1993)
Science 262, 1982-1988
   Abstract »    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
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Involvement of U6 snRNA in 5' splice site selection.
S Kandels-Lewis and B Seraphin (1993)
Science 262, 2035-2039
   Abstract »    PDF »
A splicing enhancer in the human fibronectin alternate ED1 exon interacts with SR proteins and stimulates U2 snRNP binding..
A Lavigueur, H La Branche, A R Kornblihtt, and B Chabot (1993)
Genes & Dev. 7, 2405-2417
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Four yeast spliceosomal proteins (PRP5, PRP9, PRP11, and PRP21) interact to promote U2 snRNP binding to pre-mRNA..
S W Ruby, T H Chang, and J Abelson (1993)
Genes & Dev. 7, 1909-1925
   Abstract »    PDF »
Conformational changes of U6 RNA during the spliceosome cycle: an intramolecular helix is essential both for initiating the U4-U6 interaction and for the first step of slicing..
T Wolff and A Bindereif (1993)
Genes & Dev. 7, 1377-1389
   Abstract »    PDF »
A functional association between the 5' and 3' splice site is established in the earliest prespliceosome complex (E) in mammals..
S Michaud and R Reed (1993)
Genes & Dev. 7, 1008-1020
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Movement of the guide sequence during RNA catalysis by a group I ribozyme.
J. Wang, W. Downs, and T. Cech (1993)
Science 260, 504-508
   Abstract »    PDF »
Association with terminal exons in pre-mRNAs: a new role for the U1 snRNP?.
K M Wassarman and J A Steitz (1993)
Genes & Dev. 7, 647-659
   Abstract »    PDF »
Cloning and characterization of PSF, a novel pre-mRNA splicing factor..
J G Patton, E B Porro, J Galceran, P Tempst, and B Nadal-Ginard (1993)
Genes & Dev. 7, 393-406
   Abstract »    PDF »
Splicer RNAs.
J. Steitz (1993)
Science 259, 1240
   PDF »
Site-specific cross-linking of mammalian U5 snRNP to the 5' splice site before the first step of pre-mRNA splicing..
J R Wyatt, E J Sontheimer, and J A Steitz (1992)
Genes & Dev. 6, 2542-2553
   Abstract »    PDF »
Localization and Stability of Introns Spliced from the Pem Homeobox Gene.
J. Q. Clement, S. Maiti, and M. F. Wilkinson (2001)
J. Biol. Chem. 276, 16919-16930
   Abstract »    Full Text »    PDF »
In Vitro RNA Synthesis from Exogenous Dengue Viral RNA Templates Requires Long Range Interactions between 5'- and 3'-Terminal Regions That Influence RNA Structure.
S. You, B. Falgout, L. Markoff, and R. Padmanabhan (2001)
J. Biol. Chem. 276, 15581-15591
   Abstract »    Full Text »    PDF »


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