An Early Hierarchic Role of U1 Small Nuclear Ribonucleoprotein in Spliceosome Assembly
- Creators
- Ruby, Stephanie W.
- Abelson, John
Abstract
Splicing of nuclear precursor messenger RNA (pre-mRNA) occurs on a large ribonucleoprotein complex, the spliceosome. Several small nuclear ribonucleoproteins (snRNP's) are subunits of this complex that assembles on the pre-mRNA. Although the U1 snRNP is known to recognize the 5' splice site, its roles in spliceosome formation and splice site alignment have been unclear. A new affinity purification method for the spliceosome is described which has provided insight into the very early stages of spliceosome formation in a yeast in vitro splicing system. Surprisingly, the U1 snRNP initially recognizes sequences at or near both splice junctions in the intron. This interaction must occur before the other snRNP's (U2, U4, U5, and U6) can join the complex. 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.
Additional Information
© 1988 American Association for the Advancement of Science. Received 3 August 1988; accepted 13 October 1988. We thank Z. Hostomsky for an early version of a solid-phase splicing assay; M. Ares (snR20), B. Patterson and C. Guthrie (snR7), P. G. Silciano and C. Guthrie (snR14 and snR19), P. Fabrizio (snR6), U. Vijayraghavan (mutant actins), and J. Szostak (rDNA) for cloned genes; O. Uhlenbeck for helpful advice on in vitro synthesis of small RNA's; S. Horvath for synthesizing oligodeoxynucleotides; R. Luhrmann for providing antibody to m3G cap; M. Aebi, T. Amatruda, G. McFarland, and U. Vijayraghavan for commenting on the manuscript; and D. Brow, C. Guthrie, A. Igel, M. Ares, E. Shuster, P. Legrain, B. Seraphin, and M. Rosbash for communicating results prior to publication. Supported by NIH grant GM32637 (J.A.) and by American Cancer Society Fellowship PF2647 (S.W.R.).Additional details
- Eprint ID
- 53128
- Resolver ID
- CaltechAUTHORS:20141223-075300403
- GM32637
- NIH
- PF2647
- American Cancer Society
- Created
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2014-12-23Created from EPrint's datestamp field
- Updated
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2021-11-10Created from EPrint's last_modified field