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Science 13 June 1997:
Vol. 276. no. 5319, pp. 1712 - 1716
DOI: 10.1126/science.276.5319.1712
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Reports

Bimolecular Exon Ligation by the Human Spliceosome

Karin Anderson, Melissa J. Moore *

Intron excision is an essential step in eukaryotic gene expression, but the molecular mechanisms by which the spliceosome accurately identifies splice sites in nuclear precursors to messenger RNAs (pre-mRNAs) are not well understood. A bimolecular assay for the second step of splicing has now revealed that exon ligation by the human spliceosome does not require covalent attachment of a 3' splice site to the branch site. Furthermore, accurate definition of the 3' splice site in this system is independent of either a covalently attached polypyrimidine tract or specific 3' exon sequences. Rather, in this system 3' splice site selection apparently occurs with a 5' rightarrow  3' directionality.

W. M. Keck Institute for Cellular Visualization, Department of Biochemistry, Brandeis University, Waltham, MA 02254, USA.
*   To whom correspondence should be addressed. E-mail: mmoore{at}binah.cc.brandeis.edu

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Y. Dou, K. L. Fox-Walsh, P. F. Baldi, and K. J. Hertel (2006)
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Characterization of sequences and mechanisms through which ISE/ISS-3 regulates FGFR2 splicing.
R. H. Hovhannisyan, C. C. Warzecha, and R. P. Carstens (2006)
Nucleic Acids Res. 34, 373-385
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Systematic Study of Sequence Motifs for RNA trans Splicing in Trypanosoma brucei.
T. N. Siegel, K. S. W. Tan, and G. A. M. Cross (2005)
Mol. Cell. Biol. 25, 9586-9594
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5' exon interactions within the human spliceosome establish a framework for exon junction complex structure and assembly.
V. L. Reichert, H. Le Hir, M. S. Jurica, and M. J. Moore (2002)
Genes & Dev. 16, 2778-2791
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Reversible inhibition of the second step of splicing suggests a possible role of zinc in the second step of splicing.
N. Shomron, H. Malca, I. Vig, and G. Ast (2002)
Nucleic Acids Res. 30, 4127-4137
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Sequences upstream of the branch site are required to form helix II between U2 and U6 snRNA in a trans-splicing reaction.
G. Ast, T. Pavelitz, and A. M. Weiner (2001)
Nucleic Acids Res. 29, 1741-1749
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Role of the 3' Splice Site in U12-Dependent Intron Splicing.
R. C. Dietrich, M. J. Peris, A. S. Seyboldt, and R. A. Padgett (2001)
Mol. Cell. Biol. 21, 1942-1952
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Switch in 3' Splice Site Recognition between Exon Definition and Splicing Catalysis Is Important for Sex-lethal Autoregulation.
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Mol. Cell. Biol. 21, 1986-1996
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An Upstream AG Determines Whether a Downstream AG Is Selected during Catalytic Step II of Splicing.
K. Chua and R. Reed (2001)
Mol. Cell. Biol. 21, 1509-1514
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Evidence for a linear search in bimolecular 3' splice site AG selection.
S. Chen, K. Anderson, and M. J. Moore (2000)
PNAS 97, 593-598
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An exonic splicing silencer in the testes-specific DNA ligase III {beta} exon.
S. L. Chew, L. Baginsky, and I. C. Eperon (2000)
Nucleic Acids Res. 28, 402-410
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Better conditions for mammalian in vitro splicing provided by acetate and glutamate as potassium counterions.
V. Reichert and M. J. Moore (2000)
Nucleic Acids Res. 28, 416-423
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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
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Evidence for the function of an exonic splicing enhancer after the first catalytic step of pre-mRNA splicing.
S. L. Chew, H.-X. Liu, A. Mayeda, and A. R. Krainer (1999)
PNAS 96, 10655-10660
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Serine-arginine (SR)-rich splicing factors have an exon-independent function in pre-mRNA splicing.
K. J. Hertel and T. Maniatis (1999)
PNAS 96, 2651-2655
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